Sphingosine 1 phosphate receptor modulators and methods of chiral synthesis

ABSTRACT

Compounds that selectively modulate the sphingosine 1 phosphate receptor are provided including compounds which modulate subtype 1 of the S1P receptor. Methods of chiral synthesis of such compounds are provided. Uses, methods of treatment or prevention and methods of preparing inventive compositions including inventive compounds are provided in connection with the treatment or prevention of diseases, malconditions, and disorders for which modulation of the sphingosine 1 phosphate receptor is medically indicated.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 13/740,669 filed Jan. 14, 2013, which is a divisional of U.S.application Ser. No. 12/946,800, filed Nov. 15, 2010 (granted—U.S. Pat.No. 8,357,706 issued Jan. 22, 2013, which claims benefit of U.S.Provisional Application No. 61/261,282, filed Nov. 13, 2009 and U.S.Provisional Application No. 61/262,474, filed Nov. 18, 2009, thedisclosures of which are incorporated herein in their entireties.

FIELD OF THE INVENTION

The invention relates to compounds which are agonists of the sphingosine1-phosphate receptor subtype 1, methods of their synthesis and methodsof their therapeutic and/or prophylactic use.

BACKGROUND

The S1P₁/EDG₁ receptor is a G-protein coupled receptor (GPCR) and is amember of the endothelial cell differentiation gene (EDG) receptorfamily. Endogenous ligands for EDG receptors include lysophospholipids,such as sphingosine-1-phosphate (S1P). Like all GPCRs, ligation of thereceptor propagates second messenger signals via activation ofG-proteins (alpha, beta and gamma).

Development of small molecule SIP′ agonists and antagonists has providedinsight into some physiological roles of the S1P₁/S1P-receptor signalingsystem. Agonism of the S1P₁ receptor perturbs lymphocyte trafficking,sequestering them in lymph nodes and other secondary lymphoid tissue.This leads to rapid and reversible lymphopenia, and is probably due toreceptor ligation on both lymphatic endothelial cells and lymphocytesthemselves (Rosen et al, Immunol. Rev., 195:160-177, 2003). A clinicallyvaluable consequence of lymphocyte sequestration is exclusion of themfrom sights of inflammation and/or auto-immune reactivity in peripheraltissues.

Agonism of S1P₁ has also been reported to promote survival ofoligodendrocyte progenitors (Miron et al, Ann. Neurol., 63:61-71, 2008).This activity, in conjunction with lymphocyte sequestration would beuseful in treating inflammatory and autoimmune conditions of the centralnervous system.

SUMMARY OF THE INVENTION

The present invention is directed to heterocyclic compounds adapted toact as agonists of S1P receptor subtype 1, S1P₁; methods of preparationand methods of use, such as in treatment of a malcondition mediated byS1P₁ activation, or when activation of S1P₁ is medically indicated.

Certain embodiments of the present invention comprise a compound havingthe structure of Formula I-R or I-S or a pharmaceutically acceptablesalt, ester, prodrug, homolog, hydrate or solvate thereof:

X can be —NR′R″ or —OR′″ and Y can be —CN, —Cl, or —CF₃.

R′ can be H, C₁₋₄ alkyl, n-hydroxy C₁₋₄ alkyl, —SO₂—R¹, or —CO—R′. R″can be H, —SO₂—R³, C₁₋₄ alkyl optionally substituted with 1 or more R²,or a ring moiety optionally substituted with R⁴ wherein such ring moietyis piperidinyl, cyclohexyl, morpholinyl, pyrrolidinyl, imidazolyl, orphenyl. R′″ can be H, C₁₋₄ alkyl, or —CO—R′. Alternatively, R′ and R″taken together with the nitrogen atom to which they are bound form a 4,5, or 6 membered saturated heterocyclic ring containing 0 or 1additional heteroatoms where such additional heteroatom is O or Nwherein such heterocycle is optionally singly or multiply substitutedwith substituents independently selected from —OH, oxo, —NH₂,n-hydroxy-C₁₋₄ alkyl, —COOH, —(CH₂)_(m)—COOH, —(CH₂)_(m)—COOR¹,—N(R¹R¹), and —(CH₂)_(m)—CO—N(R⁵R⁵).Each R′ can be independently C₁₋₄ alkyl or H and each R² can beindependently H, halo, OH, oxo, ═NH, NH₂, —COOH, F, —NHR′,—N(R⁵R⁵),—SO₂—R′, —SO₂—N(R⁵R⁵), —N(R′)—SO₂—R′, —COOR′, —OCO—R′,—CO—N(R⁵R⁵), —N(R′)—COR′, C₁₋₃ alkyl, C₁₋₃ alkoxy, and a ring moietyoptionally substituted with R⁴ wherein such ring moiety is piperazinyl,piperidinyl, morpholinyl, pyrrolidinyl, pyrazolyl, imidazolyl,benzimidazolyl, azetidinyl, cyclobutinyl, or phenyl.Each R³ can be independently R², C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or C₁₋₄alkyl optionally substituted with 1 or more R²; and each R⁴ can beindependently halo, OH, —NH₂, —NHR′, —N(R′R′), —COOH, —COOR′, —NHCO—R′.Each R⁵ can be independently C₁₋₄ alkyl or H, or alternatively two R⁵taken together with the nitrogen atom to which they are bound can form a4, 5, or 6 membered saturated heterocyclic ring containing 0 or 1additional heteroatoms where such additional heteroatom is O or Nwherein such heterocycle is optionally substituted with —OH, —NH₂,—N(R′R′), n-hydroxy C₁₋₄ alkyl, —(CH₂)_(m)—COOH, —(CH₂)_(m)—COOR¹. Eachm is independently 0, 1, 2, or 3.

In certain embodiments, a pharmaceutical composition comprising acompound of the invention and a suitable excipient is provided.

In certain embodiments, a method of use of an inventive compoundcomprising preparation of a medicament is provided.

In certain combinations, a pharmaceutical combination comprising acompound of the invention and a second medicament is provided. Invarious embodiments the second medicament is medically indicated for thetreatment of multiple sclerosis, transplant rejection, acute respiratorydistress syndrome or adult respiratory distress syndrome.

In certain embodiments, a method of activation or agonism of asphingosine-1-phosphate receptor subtype 1 comprising contacting thereceptor subtype 1 with a compound of claim 1 is provided. In variousembodiments, the compound of claim 1 activates or agonizes thesphingosine-1-phosphate receptor subtype 1 to a greater degree than thecompound activates or agonizes a sphingosin-1-phosphate receptor subtype3.

In certain embodiments a method of treatment of a malcondition in apatient for which activation or agonism of an S1P₁ receptor is medicallyindicated, is provided. In various embodiment, selective activation oragonism of an S1P₁ receptor, such as with respect to an S1P₃ receptor,is medically indicated. In various embodiments, the malconditioncomprises multiple sclerosis, transplant rejection, or acute respiratorydistress syndrome.

In certain embodiments, a method is provided for chiral synthesis ofcertain compounds including compounds of the invention. In certain otherembodiments the invention provides certain intermediate compoundsassociated with such methods of chiral synthesis.

DETAILED DESCRIPTION OF THE INVENTION

Certain embodiments of the present invention comprise a compound havingthe structure of Formula I-R or I-S or a pharmaceutically acceptablesalt, ester, prodrug, homolog, hydrate or solvate thereof:

X can be —NR′R″ or —OR′″ and Y can be —CN, —Cl, or —CF₃. R′ can be H,C₁₋₄ alkyl, n-hydroxy C₁₋₄ alkyl, —SO₂—R¹, or —CO—R′. R″ can be H,—SO₂—R³, C₁₋₄ alkyl optionally substituted with 1 or more R², or a ringmoiety optionally substituted with R⁴ wherein such ring moiety ispiperidinyl, cyclohexyl, morpholinyl, pyrrolidinyl, imidazolyl, orphenyl. R′″ can be H, C₁₋₄ alkyl, or —CO—R¹. Alternatively, R′ and R″taken together with the nitrogen atom to which they are bound form a 4,5, or 6 membered saturated heterocyclic ring containing 0 or 1additional heteroatoms where such additional heteroatom is O or Nwherein such heterocycle is optionally singly or multiply substitutedwith substituents independently selected from —OH, oxo, —NH₂,n-hydroxy-C₁₋₄ alkyl, —COOH, —(CH₂)_(m)—COOH, —(CH₂)_(m)—COOR¹,—N(R¹R¹), and —(CH₂)_(m)—CO—N(R⁵R⁵).

Each R¹ can be independently C₁₋₄ alkyl or H and each R² can beindependently H, halo, OH, oxo, ═NH, NH₂, —COOH, F, —NHR′,—N(R⁵R⁵),—SO₂— R′, —SO₂—N(R⁵R⁵), —N(R¹)—SO₂—R¹, —COOR¹, —OCO—R¹,—CO—N(R⁵R⁵), —N(R¹)—COR¹, C₁₋₃ alkyl, C₁₋₃ alkoxy, and a ring moietyoptionally substituted with R⁴ wherein such ring moiety is piperazinyl,piperidinyl, morpholinyl, pyrrolidinyl, pyrazolyl, thiazolyl,imidazolyl, benzimidazolyl, azetidinyl, cyclobutinyl, or phenyl.

Each R³ can be independently R², C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or C₁₋₄alkyl optionally substituted with 1 or more R²; and each R⁴ can beindependently halo, OH, —NH₂, —NHR¹, —N(R¹R¹), —COOH, —COOR¹, —NHCO—R¹.Each R⁵ can be independently C₁₋₄ alkyl or H, or alternatively two R⁵taken together with the nitrogen atom to which they are bound can form a4, 5, or 6 membered saturated heterocyclic ring containing 0 or 1additional heteroatoms where such additional heteroatom is O or Nwherein such heterocycle is optionally substituted with —OH, —NH₂,—N(R′R′), n-hydroxy C₁₋₄ alkyl, —(CH₂)_(m)—COOH, —(CH₂)_(m)—COOR¹. Eachm is independently 0, 1, 2, or 3.

In certain embodiments, the compounds of the invention have thestructure of Formula I-R or a pharmaceutically acceptable salt, ester,prodrug, homolog, hydrate or solvate thereof. In other embodiments, thecompounds of the invention have the structure of Formula I-S or apharmaceutically acceptable salt, ester, prodrug, homolog, hydrate orsolvate thereof.

In certain embodiments the invention provides compounds which aresubstantially enantiomerically pure.

In certain embodiments the invention provides compounds which have anEC₅₀ as an agonist of the wild type S1P receptor subtype 1 which is atleast ten times smaller than the EC₅₀ of such compound as an agonist ofa mutant S1P receptor subtype 1 having a single mutation with respect towild type S1P receptor subtype 1 such that the 101^(st) amino acidresidue is changed from asparagine to alanine.

In certain embodiments the invention provides compounds which have anEC₅₀ as an agonist of the wild type S1P receptor subtype 1 which is atleast twenty times smaller than the EC₅₀ of such compound as an agonistof a mutant S1P receptor subtype 1 having a single mutation with respectto wild type S1P receptor subtype 1 such that the 101^(st) amino acidresidue is changed from asparagine to alanine.

In certain embodiments the invention provides compounds which have atherapeutic index of at least 5 as measured in rats following 5 or 14days of dosing of rats with the compound where the therapeutic index iscalculated as a ratio of (i) the highest dose of such compound whichachieves less than or equal to a ten percent increase in the ratio oflung to terminal body weight at the conclusion of such 5 or 14 days ofdosing, to (ii) the dose of such compound achieving 50% lymphopenia inrats. In certain embodiments, such therapeutic index is at least 10 andin certain embodiments the therapeutic index is at least 20. In certainembodiments, the therapeutic index for a compound is at least five timesgreater than the therapeutic index for the enantiomer of such compound.

In certain embodiments the invention provides compounds which have atherapeutic index of at least 5 as measured in rats following 5 or 14days of dosing of rats with the compound where the therapeutic index iscalculated as a ratio of (i) the highest dose of such compound whichachieves less than or equal to a ten percent increase in the ratio oflung to terminal body weight at the conclusion of such 5 or 14 days ofdosing, to (ii) the dose of such compound achieving 50% lymphopenia inrats. In certain embodiments, such therapeutic index is at least 10 andin certain embodiments the therapeutic index is at least 20. In certainembodiments, the therapeutic index for a compound is greater than thetherapeutic index for the enantiomer of such compound. In certainembodiments, the therapeutic index for a compound is at least 150% ofthe therapeutic index for the enantiomer of such compound.

In certain embodiments the invention provides compounds where Y is Cl,in other embodiments the invention provides compounds where Y is CF₃ andin other embodiments the invention provides compounds where Y is CN.

In certain embodiments the invention provides compounds where X is—NR′R″, in other embodiments the invention provides compounds where X is—OR′″. In certain embodiments the invention provides compounds where Xis —OR′″. In certain embodiments the invention provides compounds whereX is —OH and in other embodiments the invention provides compounds whereX is —OCO—R¹.

In certain embodiments the invention provides compounds where R₁ is C₁₋₃alkyl; in other embodiments the invention provides compounds where R′ isH.

In certain embodiments the invention provides compounds where R′ is—COR¹; in other embodiments the invention provides compounds where R′ isSO₂—R¹. In certain embodiments the invention provides compounds where R″is H.

In certain embodiments the invention provides compounds where R″ is—SO₂—R³; in other embodiments the invention provides compounds where R″is C₁₋₄ alkyl where the C₁₋₄ alkyl is optionally substituted with 1 ormore substituents defined by R². In certain embodiments the inventionprovides compounds where R″ is —(CR^(a)R^(b))_(n)—R² and each R^(a) andeach R^(b) can be independently any of H, hydroxyl and methyl or whereR^(a) and R^(b) are bound to the same carbon they can be taken togetherto form oxo (i.e. with the carbon to which they are bound forming acarbonyl moiety). In certain such embodiments n can be 0, 1, 2, or 3 andin certain embodiments n is 2. In certain such embodiments R₂ can be—OH, —NH₂, —NHR¹, —N(R⁵R⁵), or —COOH.

In certain embodiments the invention provides compounds where R³ is C₁₋₄alkyl optionally substituted with 1 or more R². In certain embodimentsthe invention provides compounds where R² is OH; in other embodimentsthe invention provides compounds where R² is C₁₋₃ alkoxy. In certainembodiments the invention provides compounds where R³ is (CH₂)₂—OR¹.

In certain embodiments the invention provides compounds where Y is CNand X is —NH—SO₂—R³. In certain embodiments the invention providescompounds where R³ is —C₂H₅—N((R⁵R⁵) or —CH₂—CO—N(R⁵R⁵). In certainembodiments the invention provides compounds where Y is CN and X is—NH—CO—N(R⁵R⁵).

In certain embodiments X is —NH₂ and in certain of such embodiments Y isCN.

In certain embodiments the invention provides one or more of compounds1-55:

or any pharmaceutically acceptable salt, tautomer, stereoisomer,solvate, hydrate, or prodrug thereof. In certain of such embodiments,the invention provides a compound selected from compounds 8, 13, 29, 33,37, and 49 or any pharmaceutically acceptable salt, ester, tautomer,stereoisomer, solvate, hydrate, homolog, or prodrug thereof.

In certain embodiments, an invention compound of Formula I is providedwherein the compound has at least one chiral center and is substantiallyenantiomerically pure.

In other embodiments, a pharmaceutical composition comprising aninvention compound of Formula I and a suitable excipient is provided.

In other embodiments, a pharmaceutical combination comprising aninvention compound and a second medicament is provided. In still otherembodiments, a pharmaceutical combination comprising an inventioncompound and a second medicament is provided wherein the secondmedicament is medically indicated for the treatment of multiplesclerosis, transplant rejection, or adult respiratory distress syndrome.

In certain embodiments, a method of use of an invention compound forpreparation of a medicament is provided.

In certain embodiments a method of activation or agonism of asphingosine-1-phosphate receptor subtype 1 by contacting the receptorsubtype 1 with an effective amount of an invention compound. In furtherembodiments, a method of activation or agonism of asphingosine-1-phosphate receptor subtype 1 by contacting the receptorsubtype 1 with an effective amount of an invention compound is provided,wherein the compound activates or agonizes the sphingosine-1-phosphatereceptor subtype 1 to a greater extent than the compound activates oragonizes a sphingosine-1-phosphate receptor subtype 3. In furtherembodiments, a method of activation or agonism of asphingosine-1-phosphate receptor subtype 1 by contacting the receptorsubtype 1 with an effective amount of an invention compound is provided,wherein the sphingosine-1-phosphate receptor subtype 1 is disposedwithin a living mammal.

In certain embodiments, a method is provided for treatment of amalcondition in a patient for which activation or agonism of asphingosine-1-phosphate receptor subtype 1 is medically indicated, byadministering an effective amount of an invention compound to thepatient at a frequency and for a duration of time sufficient to providea beneficial effect to the patient. In further embodiments, a method isprovided for treatment of a malcondition in a patient for whichactivation or agonism of an sphingosine-1-phosphate receptor subtype 1is medically indicated, by administering an effective amount of aninvention compound to the patient at a frequency and for a duration oftime sufficient to provide a beneficial effect to the patient, whereinselective activation or agonism of an S1P subtype 1 receptor withrespect to other subtypes of S1P receptor is medically indicated. In yetfurther embodiments, a method is provided for treatment of amalcondition in a patient for which activation or agonism of ansphingosine-1-phosphate receptor subtype 1 is medically indicated, byadministering an effective amount of an invention compound to thepatient at a frequency and for a duration of time sufficient to providea beneficial effect to the patient, wherein the malcondition comprisesrejection of transplanted organs or tissue; graft-versus-host diseasesbrought about by transplantation; autoimmune syndromes includingrheumatoid arthritis; acute respiratory distress syndrome; adultrespiratory distress syndrome; influenza; cancer; systemicerythematosus; Hashimoto's thyroiditis; lymphocytic thyroiditis;multiple sclerosis; myasthenia gravis; type I and II diabetes; uveitis;posterior uveitis; uveitis associated with Behcet's disease;uveomeningitis syndrome; allergic encephalomyelitis; chronic allograftvasculopathy; post-infectious autoimmune diseases including rheumaticfever and post-infectious glomerulonephritis; inflammatory andhyperproliferative skin diseases; cutaneous manifestations ofimmunologically-mediated disorders; psoriasis; atopic dermatitis;osteomyelitis; contact dermatitis; eczematous dermatitis; seborrhoeicdermatitis; lichen planus; pemphigus; bullous pemphigoid; epidermolysisbullosa; urticaria; angioedema; vasculitis; erythema; cutaneouseosinophilia; acne; alopecia areata; keratoconjunctivitis; vernalconjunctivitis; keratitis; herpetic keratitis; dystrophia epithelialiscorneae; corneal leukoma; ocular pemphigus; Mooren's ulcer; ulcerativekeratitis; scleritis; Graves' ophthalmopathy; Vogt-Koyanagi-Haradasyndrome; sarcoidosis; pollen allergies; reversible obstructive airwaydisease; bronchial asthma; allergic asthma; intrinsic asthma; extrinsicasthma; dust asthma; chronic or inveterate asthma; late asthma andairway hyper-responsiveness; bronchitis; gastric ulcers; ischemic boweldiseases; inflammatory bowel diseases; necrotizing enterocolitis;intestinal lesions associated with thermal burns; celiac diseases;proctitis; eosinophilic gastroenteritis; mastocytosis; Crohn's disease;ulcerative colitis; vascular damage caused by ischemic diseases andthrombosis; atherosclerosis; fatty heart; myocarditis; cardiacinfarction; arteriosclerosis; aortitis syndrome; cachexia due to viraldisease; vascular thrombosis; migraine; rhinitis; eczema; interstitialnephritis; IgA-induced nephropathy; Goodpasture's syndrome;hemolytic-uremic syndrome; diabetic nephropathy; glomerulosclerosis;glomerulonephritis; multiple myositis; Guillain-Barre syndrome;Meniere's disease; polyneuritis; multiple neuritis; mononeuritis;radiculopathy; hyperthyroidism; Basedow's disease; thyrotoxicosis; purered cell aplasia; aplastic anemia; hypoplastic anemia; idiopathicthrombocytopenic purpura; autoimmune hemolytic anemia; agranulocytosis;pernicious anemia; megaloblastic anemia; anerythroplasia; osteoporosis;sarcoidosis; fibroid lung; idiopathic interstitial pneumonia;dermatomyositis; leukoderma vulgaris; ichthyosis vulgaris; photoallergicsensitivity; cutaneous T cell lymphoma; polyarteritis nodosa;Huntington's chorea; Sydenham's chorea; myocardosis; scleroderma;Wegener's granuloma; Sjogren's syndrome; adiposis; eosinophilicfascitis; lesions of gingiva, periodontium, alveolar bone, substantiaossea dentis; male pattern alopecia or alopecia senilis; musculardystrophy; pyoderma; Sezary's syndrome; chronic adrenal insufficiency;Addison's disease; ischemia-reperfusion injury of organs which occursupon preservation; endotoxin shock; pseudomembranous colitis; colitiscaused by drug or radiation; ischemic acute renal insufficiency; chronicrenal insufficiency; lung cancer; malignancy of lymphoid origin; acuteor chronic lymphocytic; leukemias; lymphoma; psoriasis; inflammatorylung injury, pulmonary emphysema; cataracta; siderosis; retinitispigmentosa; senile macular degeneration; vitreal scarring; inflammatoryeye disease; corneal alkali burn; dermatitis erythema; ballousdermatitis; cement dermatitis; gingivitis; periodontitis; sepsis;pancreatitis; carcinogenesis; metastasis of carcinoma; hypobaropathy;autoimmune hepatitis; primary biliary cirrhosis; sclerosing cholangitis;partial liver resection; acute liver necrosis; cirrhosis; alcoholiccirrhosis; hepatic failure; fulminant hepatic failure; late-onsethepatic failure; “acute-on-chronic” liver failure. In yet furtherembodiments, the malcondition is one or more of rejection oftransplanted organs or tissue; graft-versus-host diseases brought aboutby transplantation; autoimmune syndromes including rheumatoid arthritis,multiple sclerosis, myasthenia gravis; pollen allergies; type Idiabetes; prevention of psoriasis; Crohn's disease; ulcerative colitis,acute respiratory distress syndrome; adult respiratory distresssyndrome; influenza; post-infectious autoimmune diseases includingrheumatic fever and post-infectious glomerulonephritis; and metastasisof carcinoma. In yet further empbodiments the malcondition is one ofinfluenza, ulcerative colitis, multiple sclerosis, transplant rejection,acute respiratory distress syndrome or adult respiratory distresssyndrome.

In certain embodiments, methods are provided for use of an inventioncompound for preparation of a medicament adapted for treatment of adisorder or a malcondition wherein activation or inhibition of asphingosine-1-phosphate receptor subtype 1 is medically indicated.

In certain embodiments the invention provides a method for the chiralsynthesis of a compound comprising a tetrahydronaphthalene moiety havinga chiral carbon in the six-membered saturated ring of thetetrahydronaphthalene moiety where the compound is enantiomericallyenriched with respect to the chiral carbon. In such embodiments, themethod of the invention provides the steps of (i) providing a compoundcomprising a tetrahydronaphthalene moiety where the ring carbon of thesix-membered saturated ring of the tetrahydronaphthalene moiety wherechiral substitution is desired is oxo substituted at such carbon; and(ii) reacting such compound with a chiral reagent to form a chiralcenter at the tetrahydronaphthalene moiety carbon previously bound tothe oxo group. In certain of such embodiments, the chiral reagent isRuCl(p-cymene)[(R,R)-Ts-DPEN] or RuCl(p-cymene)[(S,S)-Ts-DPEN].

In certain of such embodiments the compound comprising atetrahydronaphthalene moiety provided in step (i) is contacted with thechiral reagent to form in step (ii) an intermediate of Formula VI-R orVI-S:

wherein Z is —CN, —Cl, or —CF₃. In certain of such embodiments Z is —CN.

In certain embodiments the invention provides the method comprising thestep of reversing the chiral configuration of the chiral carbon in thesix-membered saturated ring of the tetrahydronaphthalene moiety that waspreviously bound to the oxo group by treating the intermediate ofFormula VI-R or VI-S with diphenylphosphoryl azide (DPPA) to form anazido tetrahydronaphthalene of Formula VII-S or VII-R:

where the azido substituent in the six-membered saturated ring of thetetrahydronaphthalene moiety replaces the hydroxy substituent of FormulaVI-R or VI-S and the resulting chiral carbon that is bound to the azidosubstituent has a reverse chiral configuration of the chiral carbon whenit was previously bound to the hydroxy substituent.

In certain embodiments the invention provides the method where Z is —CNand the method further comprises the additional steps of (a) forming asubstituted 1,2,4-oxadiazole on the tetrahydronaphthalene moiety by (a)reacting the intermediate of VII-R or VII-S with a protecting agent andthen reacting the resulting protected form of the intermediate of VII-Ror VII-S with a hydroxylamine or a hydroxylamine hydrochloride to form ahydroxyamidine at the phenyl carbon to which Z had been attached, theresulting compound of such reaction having the Formula VIII-R or VIII-S:

(b) contacting the intermediate of Formula VIII-R or VIII-S withsubstituted benzoic acid and a coupling reagent to form a compound ofFormula IX-R or XI-S:

where X is as defined above or in certain embodiments OH, N₃, NH-PG, NH₂or NR′R″; PG can be a protecting group; R′ can be H, C₁₋₄ alkyl,n-hydroxy C₁₋₄ alkyl, —SO₂—R¹, or —CO—R¹; R″ can be H, —SO₂—R³, C₁₋₄alkyl optionally substituted with 1 or more R², or a ring moietyoptionally substituted with R⁴ wherein such ring moiety is piperidinyl,cyclohexyl, morpholinyl, thiazolyl, pyrazolyl, pyrrolidinyl, imidazolyl,or phenyl; R^(a) is lower alkyl and R¹, R², R³, and R⁴ are as definedabove. In certain of such embodiments the compounds of Formula IX-R orIX-S have the structures below:

In certain of such embodiments, the coupling reagent can be a mixturecomprising hydroxybenzotriazole (HOBt) and1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC).

Protecting groups can render chemical functionality inert to specificreaction conditions and can be appended to and removed from suchfunctionality in a molecule without substantially damaging the remainderof the molecule. Practitioners in the art would be familiar withsuitable protecting groups for use in the synthetic methods of theinvention. See, e.g., Greene and Wuts, Protective Groups in OrganicSynthesis, 2^(nd) ed., John Wiley & Sons, New York, 1991.

In certain embodiments the invention provides the method where thecompound provided in step (i) is

In certain embodiments the invention provides the method where theresulting compound comprising a tetrahydronaphthalene moiety having achiral carbon in the six-membered saturated ring of thetetrahydronaphthalene moiety is enantiomerically enriched at least 90%.In certain such embodiments the resulting compound is enantiomericallyenriched at least 95%. In certain such embodiments the resultingcompound is enantiomerically enriched at least 98%. In certain suchembodiments the resulting compound is enantiomerically enriched at least99%.

In certain of such embodiments, the invention provides a method forchiral synthesis of a chiral compound comprising a tetrahydronaphthalenemoiety having a chiral carbon in the six-membered saturated ring of thetetrahydronaphthalene moiety or a chiral compound comprising anoxadiazole-tetrahydronaphthalene moiety having a chiral carbon in thesix-membered saturated ring of the tetrahydronaphthalene moiety wherethe chiral compound has an enantiomeric enrichment of at least 75%, 85%,90%, 95%, 98%, or 99%.

In certain of such embodiments, the invention provides a method forsynthesis of a chiral compound of the invention having an enantiomericenrichment of at least 75%, 85%, 90%, 95%, 98%, or 99%.

In certain embodiments, the invention provides compounds which can beintermediates in the herein described methods for chiral syntheses. Incertain such embodiments, the invention provides one or more of thefollowing intermediate compounds:

In certain of such embodiments, the invention provides a method for thesynthesis of a compound comprising a tetrahydronaphthalene moiety havinga chiral carbon in the six-membered saturated ring of thetetrahydronaphthalene moiety where the compound is enantiomericallyenriched with respect to such chiral carbon, with the method comprisinga step of providing one of such intermediate compounds.

In certain embodiments, a method for the synthesis of a compoundcomprising a tetrahydronaphthalene moiety having a chiral carbon in thesix-membered saturated ring of the tetrahydronaphthalene moiety wherethe compound is enantiomerically enriched with respect to the chiralcarbon is provided. In certain embodiments, a method comprising a stepof providing a compound of the structures described herein is provided.

In certain embodiments, the invention provides a method for thesynthesis of a compound of the Formula IX-R or XI-S:

wherein X is as defined herein, with the method comprising a step ofproviding one of the intermediate compounds described above. In certainof such embodiments the invention provides a method for the synthesis ofa compound of the invention.

In certain embodiments the invention provides a method for the chiralsynthesis of the structure of Formula IX-R or IX-S or a pharmaceuticallyacceptable salt, ester, prodrug, homolog, hydrate or solvate thereof:

where X is defined as above and where the compound is enantiomericallyenriched with respect to the chiral carbon. In such embodiments, themethod of the invention provides the steps of

-   -   (i) providing the compound

-   -    and    -   (ii) reacting such compound with a chiral reagent        RuCl(p-cymene)[(R,R)-Ts-DPEN] or RuCl(p-cymene)[(S,S)-Ts-DPEN];        and    -   (iii) forming a chiral center at the tetrahydronaphthalene        moiety carbon previously bound to the oxo group.

Additional steps for the preparation of such compounds can be adaptedfrom the synthetic methods disclosed herein including recrystallizationand other processes for purification.

As used in the specification and the appended claims, the singular forms“a,” “an” and “the” include plural referents unless the context clearlydictates otherwise.

As used herein, “individual” (as in the subject of the treatment) meansboth mammals and non-mammals. Mammals include, for example, humans;non-human primates, e.g., apes and monkeys; cattle; horses; sheep; andgoats. Non-mammals include, for example, fish and birds.

The term “S1P₁” as used herein refers to subtype 1 of asphingosine-1-phosphate receptor, while other sphingosine-1-phosphatereceptor subtypes are referred to in a corresponding manner, forexample, sphingosine-1-phosphate receptor subtype 3 is referred to as“S1P₃”.

A “receptor”, as is well known in the art, is a biomolecular entityusually comprising a protein that specifically binds a structural classof ligands or a single native ligand in a living organism, the bindingof which causes the receptor to transduce the binding signal intoanother kind of biological action, such as signaling a cell that abinding event has occurred, which causes the cell to alter its functionin some manner. An example of transduction is receptor binding of aligand causing alteration of the activity of a “G-protein” in thecytoplasm of a living cell. Any molecule, naturally occurring or not,that binds to a receptor and activates it for signal transduction, isreferred to as an “agonist” or “activator.” Any molecule, naturallyoccurring or not, that binds to a receptor, but does not cause signaltransduction to occur, and which can block the binding of an agonist andits consequent signal transduction, is referred to as an “antagonist.”

An “S1P₁ compound” or “S1P₁ agonist” or “S1P₁ activator” or “S1P₁inhibitor” or “S1P₁ antagonist” as the terms are used herein refer tocompounds that interact in some way with the S1P receptor subtype 1.They can be agonist or activators, or they can be antagonists orinhibitors. An “S1P₁ compound” of the invention can be selective foraction on subtype 1 of the S1P receptor family; for example a compoundof the invention can act at a lower concentration on subtype 1 of theS1P receptor family than on other subtypes of the S1P receptor family;more specifically, an “S1P₁ compound” of the invention can selectivelyact on subtype 1 receptors compared to its action on subtype 3, or“S1P₃” receptors.

In certain embodiments, compounds of the invention are orthostaticagonists. In certain other embodiments, compounds of the invention areallosteric agonists. Receptor agonists may be classified as eitherorthosteric or allosteric. An orthosteric agonist binds to a site in thereceptor that significantly overlaps with the binding of the naturalligand and replicates the key interactions of the natural ligand withthe receptor. An orthosteric agonist will activate the receptor by amolecular mechanism similar to that of the natural ligand, will becompetitive for the natural ligand, and will be competitivelyantagonized by pharmacological agents that are competitive antagonistsfor the natural ligand. An allosteric agonist binds to a site in thereceptor that makes some significant interactions that are partly orwholly non-overlapping with the natural ligand. Allosteric agonists aretrue agonists and not allosteric potentiators. Consequently, theyactivate receptor signaling alone and without a requirement for asub-maximal concentration of the natural ligand. Allosteric agonists maybe identified when an antagonist known to be competitive for theorthosteric ligand shows non-competitive antagonism. The allostericagonist site can also be mapped by receptor mutagenesis. Theintroduction of single point mutations in receptors that retain receptoractivation by allosteric agonist, while diminishing or abolishingsignaling induced by orthosteric agonist or vice versa provide formalevidence for differences in binding interactions. Orthosteric agonistsmay destabilize GPCR structure and conformation, while allostericagonists may either stabilize or destabilize GPCR structure andconformation. Allosteric agonists, by virtue of their differentinteractions with receptor, may be pharmaceutically useful because theallosteric site may confer additional opportunities for agonist potencyand selectivity within a related family of receptor subtypes that sharea similar orthosteric ligand. In addition, the allosteric site mayrequire very different physical and chemical properties of an agonistcompared to the orthosteric ligand. These chemico-physical properties,which include hydrophobicity, aromaticity, charge distribution andsolubility, may also provide advantages in generating agonists ofvarying pharmacokinetic, oral bioavailability, distributional andmetabolism profiles that facilitate the development of effectivepharmaceutical substances.

“Substantially” as the term is used herein means completely or almostcompletely; for example, a composition that is “substantially free” of acomponent either has none of the component or contains such a traceamount that any relevant functional property of the composition isunaffected by the presence of the trace amount, or a compound is“substantially pure” is there are only negligible traces of impuritiespresent.

Substantially enantiomerically pure means a level of enantiomericenrichment of one enantiomer with respect to the other enantiomer of atleast 90%, 95%, 98%, 99%, 99.5% or 99.9%.

“Treating” or “treatment” within the meaning herein refers to analleviation of symptoms associated with a disorder or disease, orinhibition of further progression or worsening of those symptoms, orprevention or prophylaxis of the disease or disorder.

The expression “effective amount”, when used to describe use of acompound of the invention in providing therapy to a patient sufferingfrom a disorder or malcondition mediated by a sphingosine-1-phospatereceptor of subtype 1 refers to the amount of a compound of theinvention that is effective to bind to as an agonist or as an antagonista S1P₁ receptor in the individual's tissues, wherein the S1P₁ isimplicated in the disorder, wherein such binding occurs to an extentsufficient to produce a beneficial therapeutic effect on the patient.Similarly, as used herein, an “effective amount” or a “therapeuticallyeffective amount” of a compound of the invention refers to an amount ofthe compound that alleviates, in whole or in part, symptoms associatedwith the disorder or condition, or halts or slows further progression orworsening of those symptoms, or prevents or provides prophylaxis for thedisorder or condition. In particular, a “therapeutically effectiveamount” refers to an amount effective, at dosages and for periods oftime necessary, to achieve the desired therapeutic result by acting asan agonist of sphingosine-1-phosphate receptor subtype 1 (S1P₁)activity. A therapeutically effective amount is also one in which anytoxic or detrimental effects of compounds of the invention areoutweighed by the therapeutically beneficial effects. For example, inthe context of treating a malcondition mediated by activation of S1P₁, atherapeutically effective amount of an S1P₁ agonist of the invention isan amount sufficient to control the malcondition, to mitigate theprogress of the malcondition, or to relieve the symptoms of themalcondition. Examples of malconditions that can be so treated includemultiple sclerosis, transplant rejection, adult respiratory distresssyndrome.

Diseases, disorders and conditions which may be treated by compounds ofthe invention include rejection of transplanted organs or tissue;graft-versus-host diseases brought about by transplantation; autoimmunesyndromes including rheumatoid arthritis; acute respiratory distresssyndrome; adult respiratory distress syndrome; influenza; cancer;systemic erythematosus; Hashimoto's thyroiditis; lymphocyticthyroiditis; multiple sclerosis; myasthenia gravis; type I and IIdiabetes; uveitis; posterior uveitis; uveitis associated with Behcet'sdisease; uveomeningitis syndrome; allergic encephalomyelitis; chronicallograft vasculopathy; post-infectious autoimmune diseases includingrheumatic fever and post-infectious glomerulonephritis; inflammatory andhyperproliferative skin diseases; cutaneous manifestations ofimmunologically-mediated disorders; psoriasis; atopic dermatitis;osteomyelitis; contact dermatitis; eczematous dermatitis; seborrhoeicdermatitis; lichen planus; pemphigus; bullous pemphigoid; epidermolysisbullosa; urticaria; angioedema; vasculitis; erythema; cutaneouseosinophilia; acne; alopecia areata; keratoconjunctivitis; vernalconjunctivitis; keratitis; herpetic keratitis; dystrophia epithelialiscorneae; corneal leukoma; ocular pemphigus; Mooren's ulcer; ulcerativekeratitis; scleritis; Graves' ophthalmopathy; Vogt-Koyanagi-Haradasyndrome; sarcoidosis; pollen allergies; reversible obstructive airwaydisease; bronchial asthma; allergic asthma; intrinsic asthma; extrinsicasthma; dust asthma; chronic or inveterate asthma; late asthma andairway hyper-responsiveness; bronchitis; gastric ulcers; ischemic boweldiseases; inflammatory bowel diseases; necrotizing enterocolitis;intestinal lesions associated with thermal burns; celiac diseases;proctitis; eosinophilic gastroenteritis; mastocytosis; Crohn's disease;ulcerative colitis; vascular damage caused by ischemic diseases andthrombosis; atherosclerosis; fatty heart; myocarditis; cardiacinfarction; arteriosclerosis; aortitis syndrome; cachexia due to viraldisease; vascular thrombosis; migraine; rhinitis; eczema; interstitialnephritis; IgA-induced nephropathy; Goodpasture's syndrome;hemolytic-uremic syndrome; diabetic nephropathy; glomerulosclerosis;glomerulonephritis; multiple myositis; Guillain-Barre syndrome;Meniere's disease; polyneuritis; multiple neuritis; mononeuritis;radiculopathy; hyperthyroidism; Basedow's disease; thyrotoxicosis; purered cell aplasia; aplastic anemia; hypoplastic anemia; idiopathicthrombocytopenic purpura; autoimmune hemolytic anemia; agranulocytosis;pernicious anemia; megaloblastic anemia; anerythroplasia; osteoporosis;sarcoidosis; fibroid lung; idiopathic interstitial pneumonia;dermatomyositis; leukoderma vulgaris; ichthyosis vulgaris; photoallergicsensitivity; cutaneous T cell lymphoma; polyarteritis nodosa;Huntington's chorea; Sydenham's chorea; myocardosis; scleroderma;Wegener's granuloma; Sjogren's syndrome; adiposis; eosinophilicfascitis; lesions of gingiva, periodontium, alveolar bone, substantiaossea dentis; male pattern alopecia or alopecia senilis; musculardystrophy; pyoderma; Sezary's syndrome; chronic adrenal insufficiency;Addison's disease; ischemia-reperfusion injury of organs which occursupon preservation; endotoxin shock; pseudomembranous colitis; colitiscaused by drug or radiation; ischemic acute renal insufficiency; chronicrenal insufficiency; lung cancer; malignancy of lymphoid origin; acuteor chronic lymphocytic; leukemias; lymphoma; psoriasis; inflammatorylung injury, pulmonary emphysema; cataracta; siderosis; retinitispigmentosa; senile macular degeneration; vitreal scarring; inflammatoryeye disease; corneal alkali burn; dermatitis erythema; ballousdermatitis; cement dermatitis; gingivitis; periodontitis; sepsis;pancreatitis; carcinogenesis; metastasis of carcinoma; hypobaropathy;autoimmune hepatitis; primary biliary cirrhosis; sclerosing cholangitis;partial liver resection; acute liver necrosis; cirrhosis; alcoholiccirrhosis; hepatic failure; fulminant hepatic failure; late-onsethepatic failure; “acute-on-chronic” liver failure. Particularlypreferred diseases and conditions which may be treated with compounds ofthe invention comprise the group consisting of rejection of transplantedorgans or tissue; graft-versus-host diseases brought about bytransplantation; autoimmune syndromes including rheumatoid arthritis,multiple sclerosis, myasthenia gravis; pollen allergies; type Idiabetes; prevention of psoriasis; Crohn's disease; ulcerative colitis,acute respiratory distress syndrome; adult respiratory distresssyndrome; influenza; post-infectious autoimmune diseases includingrheumatic fever and post-infectious glomerulonephritis; and metastasisof carcinoma.

Furthermore, compounds of Formula I-R or I-S are also useful, incombination with one or several immunosuppressant agents, for thetreatment of diseases, disorders and conditions associated with anactivated immune system and selected from the list as above-mentioned.According to a preferred embodiment of the invention, saidimmunosuppressant agent is selected from the group comprising orconsisting of cyclosporin, daclizumab, basiliximab, everolimus,tacrolimus (FK506), azathiopirene, leflunomide, 15-deoxyspergualin, orother immunosuppressant drugs

All chiral, diastereomeric, racemic forms of a structure are intended,unless a particular stereochemistry or isomeric form is specificallyindicated. Compounds used in the present invention can include enrichedor resolved optical isomers at any or all asymmetric atoms as areapparent from the depictions, at any degree of enrichment. Both racemicand diastereomeric mixtures, as well as the individual optical isomerscan be synthesized so as to be substantially free of their enantiomericor diastereomeric partners, and these are all within the scope ofcertain embodiments of the invention.

The isomers resulting from the presence of a chiral center comprise apair of non-superimposable isomers that are called “enantiomers.” Singleenantiomers of a pure compound are optically active, i.e., they arecapable of rotating the plane of plane polarized light. Singleenantiomers are designated according to the Cahn-Ingold-Prelog system.Once the priority ranking of the four groups is determined, the moleculeis oriented so that the lowest ranking group is pointed away from theviewer. Then, if the descending rank order of the other groups proceedsclockwise, the molecule is designated (R) and if the descending rank ofthe other groups proceeds counterclockwise, the molecule is designated(S). In the examples, the Cahn-Ingold-Prelog ranking is A>B>C>D. Thelowest ranking atom, D is oriented away from the viewer.

“Isolated optical isomer” means a compound which has been substantiallypurified from the corresponding optical isomer(s) of the same formula.Preferably, the isolated isomer is at least about 80%, more preferablyat least 90% pure, even more preferably at least 98% pure, mostpreferably at least about 99% pure, by weight.

Rotational Isomerism

It is understood that due to chemical properties (i.e., resonancelending some double bond character to the C—N bond) of restrictedrotation about the amide bond linkage (as illustrated below) it ispossible to observe separate rotamer species and even, under somecircumstances, to isolate such species, example shown below. It isfurther understood that certain structural elements, including stericbulk or substituents on the amide nitrogen, may enhance the stability ofa rotamer to the extent that a compound may be isolated as, and existindefinitely, as a single stable rotamer. The present inventiontherefore includes any possible stable rotamers of compounds of theinvention which are biologically active in the treatment of a disease,disorder or condition for which a compound of the invention may beeffective as described herein.

Regioisomerism

The preferred compounds of the present invention have a particularspatial arrangement of substituents on the aromatic rings, which isrelated to the structure activity relationship demonstrated by thecompound class. Often such substitution arrangement is denoted by anumbering system; however, numbering systems are often not consistentbetween different ring systems. In six-membered aromatic systems, thespatial arrangements are specified by the common nomenclature “para” for1,4-substitution, “meta” for 1,3-substitution and “ortho” for1,2-substitution as shown below.

All structures encompassed within a claim are “chemically feasible”, bywhich is meant that the structure depicted by any combination orsubcombination of optional substituents meant to be recited by the claimis physically capable of existence with at least some stability as canbe determined by the laws of structural chemistry and byexperimentation. Structures that are not chemically feasible are notwithin a claimed set of compounds.

In general, “substituted” refers to an organic group as defined hereinin which one or more bonds to a hydrogen atom contained therein arereplaced by one or more bonds to a non-hydrogen atom such as, but notlimited to, a halogen (i.e., F, Cl, Br, and I); an oxygen atom in groupssuch as hydroxyl groups, alkoxy groups, aryloxy groups, aralkyloxygroups, oxo(carbonyl) groups, carboxyl groups including carboxylicacids, carboxylates, and carboyxlate esters; a sulfur atom in groupssuch as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups,sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atomin groups such as amines, hydroxylamines, nitriles, nitro groups,N-oxides, hydrazides, azides, and enamines; and other heteroatoms invarious other groups. Non-limiting examples of substituents that can bebonded to a substituted carbon (or other) atom include F, Cl, Br, I,OR′, OC(O)N(R′)₂, CN, CF₃, OCF₃, R′, O, S, C(O), S(O), methylenedioxy,ethylenedioxy, N(R′)₂, SR′, SOR′, SO₂R′, SO₂N(R′)₂, SO₃R′, C(O)R′,C(O)C(O)R′, C(O)CH₂C(O)R′, C(S)R′, C(O)OR′, OC(O)R′, C(O)N(R′)₂,OC(O)N(R′)₂, C(S)N(R′)₂, (CH₂)₀₋₂NHC(O)R′, (CH₂)₀₋₂N(R′)N(R)₂,N(R′)N(R′)C(O)R′, N(R′)N(R′)C(O)OR′, N(R′)N(R′)CON(R′)₂, N(R′)SO₂R′,N(R′)SO₂N(R′)₂, N(R)C(O)OR′, N(R′)C(O)R′, N(R′)C(S)R′, N(R′)C(O)N(R′)₂,N(R′)C(S)N(R′)₂, N(COR′)COR′, N(OR′)R′, C(═NH)N(R′)₂, C(O)N(OR′)R′, orC(═NOR′)R′ wherein R′ can be hydrogen or a carbon-based moiety, andwherein the carbon-based moiety can itself be further substituted.

Substituted alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl groupsas well as other substituted groups also include groups in which one ormore bonds to a hydrogen atom are replaced by one or more bonds,including double or triple bonds, to a carbon atom, or to a heteroatomsuch as, but not limited to, oxygen in carbonyl (oxo), carboxyl, ester,amide, imide, urethane, and urea groups; and nitrogen in imines,hydroxyimines, oximes, hydrazones, amidines, guanidines, and nitriles.The substituents of the substituted groups can further be substitutedwith alkyl, alkenyl, cycloalkyl, aryl, heteroaryl, and alkynyl groups asdefined herein, which can themselves be further substituted. Forexample, a C₁₋₄ alkyl group can be substituted with an amide, and theamide can further be substituted with another C₁₋₄ alkyl, which canfurther be substituted.

Substituted ring groups such as substituted aryl, heterocyclyl andheteroaryl groups also include rings and fused ring systems in which abond to a hydrogen atom is replaced with a bond to a carbon atom.Therefore, substituted aryl, heterocyclyl and heteroaryl groups can alsobe substituted with alkyl, alkenyl, cycloalkyl, aryl, heteroaryl, andalkynyl groups as defined herein, which can themselves be furthersubstituted.

The term “heteroatoms” as used herein refers to non-carbon andnon-hydrogen atoms, capable of forming covalent bonds with carbon, andis not otherwise limited. Typical heteroatoms are N, O, and S. Whensulfur (S) is referred to, it is understood that the sulfur can be inany of the oxidation states in which it is found, thus includingsulfoxides (R—S(O)—R′) and sulfones (R—S(O)₂—R′), unless the oxidationstate is specified; thus, the term “sulfone” encompasses only thesulfone form of sulfur; the term “sulfide” encompasses only the sulfide(R—S—R′) form of sulfur. When the phrases such as “heteroatoms selectedfrom the group consisting of O, NH, NR′ and S,” or “[variable] is O, S .. . ” are used, they are understood to encompass all of the sulfide,sulfoxide and sulfone oxidation states of sulfur.

Alkyl groups include straight chain and branched alkyl groups andcycloalkyl groups having from 1 to about 20 carbon atoms (C₁₋₂₀ alkyl),and typically from 1 to 12 carbons (C₁₋₁₂ alkyl) or, in someembodiments, from 1 to 8 carbon atoms (C₁₋₈ alkyl) or, in someembodiments, from 1 to 4 carbon atoms (C₁₋₄ alkyl) or, in someembodiments, from 1 to 3 carbon atoms (C₁₋₃ alkyl). Examples of straightchain alkyl groups include, but are not limited to methyl, ethyl,n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups.Examples of branched alkyl groups include, but are not limited to,isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and2,2-dimethylpropyl groups. Representative substituted alkyl groups canbe substituted one or more times with any of the groups listed above,for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, andhalogen groups. The group “n-hydroxy C₁₋₄ alkyl” represents a C₁₋₄ alkylsubstituted with a terminal hydroxy group.

Cycloalkyl groups are alkyl groups forming a ring structure, which canbe substituted or unsubstituted. Examples of cycloalkyl include, but arenot limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkylgroup has 3 to 8 ring members, whereas in other embodiments the numberof ring carbon atoms range from 3 to 5, 3 to 6, or 3 to 7. Cycloalkylgroups further include polycyclic cycloalkyl groups such as, but notlimited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, andcarenyl groups, and fused rings such as, but not limited to, decalinyl,and the like. Cycloalkyl groups also include rings that are substitutedwith straight or branched chain alkyl groups as defined above.Representative substituted cycloalkyl groups can be mono-substituted orsubstituted more than once, such as, but not limited to, 2,2-, 2,3-,2,4-2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- ortri-substituted norbornyl or cycloheptyl groups, which can besubstituted with, for example, amino, hydroxy, cyano, carboxy, nitro,thio, alkoxy, and halogen groups.

The terms “carbocyclic” and “carbocycle” denote a ring structure whereinthe atoms of the ring are carbon. In some embodiments, the carbocyclehas 3 to 8 ring members, whereas in other embodiments the number of ringcarbon atoms is 4, 5, 6, or 7. Unless specifically indicated to thecontrary, the carbocyclic ring can be substituted with as many as Nsubstituents wherein N is the size of the carbocyclic ring with forexample, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, andhalogen groups.

(Cycloalkyl)alkyl groups, also denoted cycloalkylalkyl, are alkyl groupsas defined above in which a hydrogen or carbon bond of the alkyl groupis replaced with a bond to a cycloalkyl group as defined above.

Alkenyl groups include straight and branched chain and cyclic alkylgroups as defined above, except that at least one double bond existsbetween two carbon atoms. Thus, alkenyl groups have from 2 to about 20carbon atoms, and typically from 2 to 12 carbons or, in someembodiments, from 2 to 8 carbon atoms. Examples include, but are notlimited to —CH═CH(CH₃), —CH═C(CH₃)₂, —C(CH₃)═CH₂, —C(CH₃)═CH(CH₃),—C(CH₂CH₃)═CH₂, vinyl, cyclohexenyl, cyclopentenyl, cyclohexadienyl,butadienyl, pentadienyl, and hexadienyl among others.

The term “cycloalkenyl” alone or in combination denotes a cyclic alkenylgroup wherein at least one double bond is present in the ring structure.Cycloalkenyl groups include cycloalkyl groups having at least one doublebond between two adjacent carbon atoms. Thus for example, cycloalkenylgroups include but are not limited to cyclohexenyl, cyclopentenyl, andcyclohexadienyl groups.

(Cycloalkenyl)alkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of the alkyl group is replaced with a bond to acycloalkenyl group as defined above.

Alkynyl groups include straight and branched chain alkyl groups, exceptthat at least one triple bond exists between two carbon atoms. Thus,alkynyl groups have from 2 to about 20 carbon atoms, and typically from2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms.Examples include, but are not limited to —C≡CH, —C≡C(CH₃), —C≡C(CH₂CH₃),—CH₂C≡CH, —CH₂C≡C(CH₃), and —CH₂C≡C(CH₂CH₃), among others.

Aryl groups are cyclic aromatic hydrocarbons that do not containheteroatoms. Thus aryl groups include, but are not limited to, phenyl,azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl,triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl,anthracenyl, and naphthyl groups. In some embodiments, aryl groupscontain 6-14 carbons in the ring portions of the groups. The phrase“aryl groups” includes groups containing fused rings, such as fusedaromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, andthe like), and also includes substituted aryl groups that have othergroups, including but not limited to alkyl, halo, amino, hydroxy, cyano,carboxy, nitro, thio, or alkoxy groups, bonded to one of the ring atoms.Representative substituted aryl groups can be mono-substituted orsubstituted more than once, such as, but not limited to, 2-, 3-, 4-, 5-,or 6-substituted phenyl or naphthyl groups, which can be substitutedwith groups including but not limited to those listed above.

Aralkyl groups are alkyl groups as defined above in which a hydrogen orcarbon bond of an alkyl group is replaced with a bond to an aryl groupas defined above. Representative aralkyl groups include benzyl andphenylethyl groups and fused (cycloalkylaryl)alkyl groups such as4-ethyl-indanyl. The aryl moiety or the alkyl moiety or both areoptionally substituted with other groups, including but not limited toalkyl, halo, amino, hydroxy, cyano, carboxy, nitro, thio, or alkoxygroups. Aralkenyl group are alkenyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to anaryl group as defined above.

Heterocyclyl groups include aromatic and non-aromatic ring compounds(heterocyclic rings) containing 3 or more ring members, of which one ormore is a heteroatom such as, but not limited to, N, O, S, or P. In someembodiments, heterocyclyl groups include 3 to 20 ring members, whereasother such groups have 3 to 15 ring members. At least one ring containsa heteroatom, but every ring in a polycyclic system need not contain aheteroatom. For example, a dioxolanyl ring and a benzdioxolanyl ringsystem (methylenedioxyphenyl ring system) are both heterocyclyl groupswithin the meaning herein. A heterocyclyl group designated as aC₂-heterocyclyl can be a 5-membered ring with two carbon atoms and threeheteroatoms, a 6-membered ring with two carbon atoms and fourheteroatoms and so forth. Likewise a C₄-heterocyclyl can be a 5-memberedring with one heteroatom, a 6-membered ring with two heteroatoms, and soforth. The number of carbon atoms plus the number of heteroatoms sums upto equal the total number of ring atoms. A saturated heterocyclic ringrefers to a heterocyclic ring containing no unsaturated carbon atoms.

The phrase “heterocyclyl group” includes fused ring species includingthose having fused aromatic and non-aromatic groups. The phrase alsoincludes polycyclic ring systems containing a heteroatom such as, butnot limited to, quinuclidyl and also includes heterocyclyl groups thathave substituents, including but not limited to alkyl, halo, amino,hydroxy, cyano, carboxy, nitro, thio, or alkoxy groups, bonded to one ofthe ring members. A heterocyclyl group as defined herein can be aheteroaryl group or a partially or completely saturated cyclic groupincluding at least one ring heteroatom. Heterocyclyl groups include, butare not limited to, pyrrolidinyl, furanyl, tetrahydrofuranyl,dioxolanyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl,thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl,dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl,azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl,tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups.Heterocyclyl groups can be substituted. Representative substitutedheterocyclyl groups can be mono-substituted or substituted more thanonce, including but not limited to, rings containing at least oneheteroatom which are mono, di, tri, tetra, penta, hexa, orhigher-substituted with substituents such as those listed above,including but not limited to alkyl, halo, amino, hydroxy, cyano,carboxy, nitro, thio, and alkoxy groups.

Heteroaryl groups are aromatic ring compounds containing 5 or more ringmembers, of which, one or more is a heteroatom such as, but not limitedto, N, O, and S. A heteroaryl group designated as a C₂-heteroaryl can bea 5-membered ring with two carbon atoms and three heteroatoms, a6-membered ring with two carbon atoms and four heteroatoms and so forth.Likewise a C₄-heteroaryl can be a 5-membered ring with one heteroatom, a6-membered ring with two heteroatoms, and so forth. The number of carbonatoms plus the number of heteroatoms sums up to equal the total numberof ring atoms. Heteroaryl groups include, but are not limited to, groupssuch as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl,benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl,azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, andquinazolinyl groups. The terms “heteroaryl” and “heteroaryl groups”include fused ring compounds such as wherein at least one ring, but notnecessarily all rings, are aromatic, including tetrahydroquinolinyl,tetrahydroisoquinolinyl, indolyl and 2,3-dihydro indolyl. The term alsoincludes heteroaryl groups that have other groups bonded to one of thering members, including but not limited to alkyl, halo, amino, hydroxy,cyano, carboxy, nitro, thio, or alkoxy groups. Representativesubstituted heteroaryl groups can be substituted one or more times withgroups such as those listed above.

Additional examples of aryl and heteroaryl groups include but are notlimited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl),N-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl,anthracenyl (1-anthracenyl, 2-anthracenyl, 3-anthracenyl), thiophenyl(2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl), indolyl, oxadiazolyl,isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl,acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl),imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl),triazolyl (1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol-4-yl,1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl),thiazolyl (2-thiazolyl, 4-thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl,3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl,5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3-pyridazinyl,4-pyridazinyl, 5-pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl,4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl(1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl,6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl(2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl,5-benzo[b]furanyl, 6-benzo[b]furanyl, 7-benzo[b]furanyl),2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl),3-(2,3-dihydro-benzo[b]furanyl), 4-(2,3-dihydro-benzo[b]furanyl),5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro-benzo[b]furanyl),7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl(2-benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl,5-benzo[b]thiophenyl, 6-benzo[b]thiophenyl, 7-benzo[b]thiophenyl),2,3-dihydro-benzo[b]thiophenyl, (2-(2,3-dihydro-benzo[b]thiophenyl),3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro-benzo[b]thiophenyl),5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro-benzo[b]thiophenyl),7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl,3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole(1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl,7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl,4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl,8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl),benzothiazolyl (1-benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl,5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl(1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl),5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-1-yl,5H-dibenz[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl,5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine-5-yl),10,11-dihydro-5H-dibenz[b,f]azepine(10,11-dihydro-5H-dibenz[b,f]azepine-1-yl,10,11-dihydro-5H-dibenz[b,f]azepine-2-yl,10,11-dihydro-5H-dibenz[b,f]azepine-3-yl,10,11-dihydro-5H-dibenz[b,f]azepine-4-yl,10,11-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like.

Heterocyclylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheterocyclyl group as defined above. Representative heterocyclyl alkylgroups include, but are not limited to, furan-2-yl methyl, furan-3-ylmethyl, pyridine-2-yl methyl (α-picolyl), pyridine-3-yl methyl(β-picolyl), pyridine-4-yl methyl (γ-picolyl), tetrahydrofuran-2-ylethyl, and indol-2-yl propyl. Heterocyclylalkyl groups can besubstituted on the heterocyclyl moiety, the alkyl moiety, or both.

Heteroarylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheteroaryl group as defined above. Heteroarylalkyl groups can besubstituted on the heteroaryl moiety, the alkyl moiety, or both.

By a “ring system” as the term is used herein is meant a moietycomprising one, two, three or more rings, which can be substituted withnon-ring groups or with other ring systems, or both, which can be fullysaturated, partially unsaturated, fully unsaturated, or aromatic, andwhen the ring system includes more than a single ring, the rings can befused, bridging, or spirocyclic. By “spirocyclic” is meant the class ofstructures wherein two rings are fused at a single tetrahedral carbonatom, as is well known in the art.

A “monocyclic, bicyclic or polycyclic, aromatic or partially aromaticring” as the term is used herein refers to a ring system including anunsaturated ring possessing 4n+2 pi electrons, or a partially reduced(hydrogenated) form thereof. The aromatic or partially aromatic ring caninclude additional fused, bridged, or spiro rings that are notthemselves aromatic or partially aromatic. For example, naphthalene andtetrahydronaphthalene are both a “monocyclic, bicyclic or polycyclic,aromatic or partially aromatic ring” within the meaning herein. Also,for example, a benzo-[2.2.2]-bicyclooctane is also a “monocyclic,bicyclic or polycyclic, aromatic or partially aromatic ring” within themeaning herein, containing a phenyl ring fused to a bridged bicyclicsystem. A fully saturated ring has no double bonds therein, and iscarbocyclic or heterocyclic depending on the presence of heteroatomswithin the meaning herein.

The term “alkoxy” refers to an oxygen atom connected to an alkyl group,including a cycloalkyl group, as are defined above. Examples of linearalkoxy groups include but are not limited to methoxy, ethoxy, n-propoxy,n-butoxy, n-pentyloxy, n-hexyloxy, and the like. Examples of branchedalkoxy include but are not limited to isopropoxy, sec-butoxy,tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclicalkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, and the like.

The terms “aryloxy” and “arylalkoxy” refer to, respectively, an arylgroup bonded to an oxygen atom and an aralkyl group bonded to the oxygenatom at the alkyl moiety. Examples include but are not limited tophenoxy, naphthyloxy, and benzyloxy.

An “acyl” group as the term is used herein refers to a group containinga carbonyl moiety wherein the group is bonded via the carbonyl carbonatom. The carbonyl carbon atom is also bonded to another carbon atom,which can be part of an alkyl, aryl, aralkyl cycloalkyl,cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl,heteroarylalkyl group or the like. In the special case wherein thecarbonyl carbon atom is bonded to a hydrogen, the group is a “formyl”group, an acyl group as the term is defined herein. An acyl group caninclude 0 to about 12-20 additional carbon atoms bonded to the carbonylgroup. An acyl group can include double or triple bonds within themeaning herein. An acryloyl group is an example of an acyl group. Anacyl group can also include heteroatoms within the meaning here. Anicotinoyl group (pyridyl-3-carbonyl) group is an example of an acylgroup within the meaning herein. Other examples include acetyl, benzoyl,phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and thelike. When the group containing the carbon atom that is bonded to thecarbonyl carbon atom contains a halogen, the group is termed a“haloacyl” group. An example is a trifluoroacetyl group.

The term “amine” includes primary, secondary, and tertiary amineshaving, e.g., the formula N(group)₃ wherein each group can independentlybe H or non-H, such as alkyl, aryl, and the like. Amines include but arenot limited to RNH₂, for example, alkylamines, arylamines,alkylarylamines; R₂NH wherein each R is independently selected, such asdialkylamines, diarylamines, aralkylamines, heterocyclylamines and thelike; and R₃N wherein each R is independently selected, such astrialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, andthe like. The term “amine” also includes ammonium ions as used herein.

An “amino” group is a substituent of the form —NH₂, —NHR, —NR₂, —NR₃ ⁺,wherein each R is independently selected, and protonated forms of each.Accordingly, any compound substituted with an amino group can be viewedas an amine.

An “ammonium” ion includes the unsubstituted ammonium ion NH₄ ⁺, butunless otherwise specified, it also includes any protonated orquaternarized forms of amines. Thus, trimethylammonium hydrochloride andtetramethylammonium chloride are both ammonium ions, and amines, withinthe meaning herein.

The term “amide” (or “amido”) includes C- and N-amide groups, i.e.,—C(O)N R′R″, and —NR′C(O)R″ groups, respectively. The R′ and R″ of theC-amide may join together to form a heterocyclic ring with the nitrogenatom. Amide groups therefore include but are not limited to carbamoylgroups (—C(O)NH₂) and formamide groups (—NHC(O)H). A “carboxamido” groupis a group of the formula C(O)NR₂, wherein R can be H, alkyl, aryl, etc.

The term “urethane” (or “carbamyl”) includes N- and O-urethane groups,i.e., —NRC(O)OR and —OC(O)NR₂ groups, respectively.

The term “sulfonamide” (or “sulfonamido”) includes S- and N-sulfonamidegroups, i.e., —SO₂NR₂ and —NRSO₂R groups, respectively. Sulfonamidegroups therefore include but are not limited to sulfamoyl groups(—SO₂NH₂).

The term “amidine” or “amidino” includes groups of the formula—C(NR)NR₂. Typically, an amidino group is —C(NH)NH₂.

The term “guanidine” or “guanidino” includes groups of the formula—NRC(NR)NR₂. Typically, a guanidino group is —NHC(NH)NH₂.

“Halo,” “halogen,” and “halide” include fluorine, chlorine, bromine andiodine.

The terms “comprising,” “including,” “having,” “composed of,” areopen-ended terms as used herein, and do not preclude the existence ofadditional elements or components. In a claim element, use of the forms“comprising,” “including,” “having,” or “composed of” means thatwhatever element is comprised, had, included, or composes is notnecessarily the only element encompassed by the subject of the clausethat contains that word.

A “salt” as is well known in the art includes an organic compound suchas a carboxylic acid, a sulfonic acid, or an amine, in ionic form, incombination with a counterion. For example, acids in their anionic formcan form salts with cations such as metal cations, for example sodium,potassium, and the like; with ammonium salts such as NH₄ ⁺ or thecations of various amines, including tetraalkyl ammonium salts such astetramethylammonium and alkyl ammonium salts such as tromethamine salts,or other cations such as trimethylsulfonium, and the like. A“pharmaceutically acceptable” or “pharmacologically acceptable” salt isa salt formed from an ion that has been approved for human consumptionand is generally non-toxic, such as a chloride salt or a sodium salt. A“zwitterion” is an internal salt such as can be formed in a moleculethat has at least two ionizable groups, one forming an anion and theother a cation, which serve to balance each other. For example, aminoacids such as glycine can exist in a zwitterionic form. A “zwitterion”is a salt within the meaning herein. The compounds of the presentinvention may take the form of salts. The term “salts” embraces additionsalts of free acids or free bases which are compounds of the invention.Salts can be “pharmaceutically-acceptable salts.” The term“pharmaceutically-acceptable salt” refers to salts which possesstoxicity profiles within a range that affords utility in pharmaceuticalapplications. Pharmaceutically unacceptable salts may nonethelesspossess properties such as high crystallinity, which have utility in thepractice of the present invention, such as for example utility inprocess of synthesis, purification or formulation of compounds of theinvention.

Suitable pharmaceutically-acceptable acid addition salts may be preparedfrom an inorganic acid or from an organic acid. Examples of inorganicacids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic,sulfuric, and phosphoric acids. Appropriate organic acids may beselected from aliphatic, cycloaliphatic, aromatic, araliphatic,heterocyclic, carboxylic and sulfonic classes of organic acids, examplesof which include formic, acetic, propionic, succinic, glycolic,gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic,fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic,4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic,sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric,salicylic, galactaric and galacturonic acid. Examples ofpharmaceutically unacceptable acid addition salts include, for example,perchlorates and tetrafluoroborates.

Suitable pharmaceutically acceptable base addition salts of compounds ofthe invention include, for example, metallic salts including alkalimetal, alkaline earth metal and transition metal salts such as, forexample, calcium, magnesium, potassium, sodium and zinc salts.Pharmaceutically acceptable base addition salts also include organicsalts made from basic amines such as, for example,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methylglucamine) and procaine. Examples ofpharmaceutically unacceptable base addition salts include lithium saltsand cyanate salts. Although pharmaceutically unacceptable salts are notgenerally useful as medicaments, such salts may be useful, for exampleas intermediates in the synthesis of compounds, for example in theirpurification by recrystallization. All of these salts may be prepared byconventional means from the corresponding compound by reacting, forexample, the appropriate acid or base with the compound. The term“pharmaceutically acceptable salts” refers to nontoxic inorganic ororganic acid and/or base addition salts, see, for example, Lit et al.,Salt Selection for Basic Drugs (1986), Int J. Pharm., 33, 201-217,incorporated by reference herein

Nonlimiting examples of potential salts of this invention include butare not limited to hydrochloride, citrate, glycolate, fumarate, malate,tartrate, mesylate, esylate, cinnamate, isethionate, sulfate, phosphate,diphosphate, nitrate, hydrobromide, hydroiodide, succinate, formate,acetate, dichloroacetate, lactate, p-toluenesulfonate, pamitate,pidolate, pamoate, salicylate, 4-aminosalicylate, benzoate, 4-acetamidobenzoate, glutamate, aspartate, glycolate, adipate, alginate, ascorbate,besylate, camphorate, camphorsulfonate, camsylate, caprate, caproate,cyclamate, laurylsulfate, edisylate, gentisate, galactarate, gluceptate,gluconate, glucuronate, oxoglutarate, hippurate, lactobionate, malonate,maleate, mandalate, napsylate, napadisylate, oxalate, oleate, sebacate,stearate, succinate, thiocyanate, undecylenate, and xinafoate.

A “hydrate” is a compound that exists in a composition with watermolecules. The composition can include water in stoichiometicquantities, such as a monohydrate or a dihydrate, or can include waterin random amounts. As the term is used herein a “hydrate” refers to asolid form, i.e., a compound in water solution, while it may behydrated, is not a hydrate as the term is used herein.

A “homolog” of a compound of the invention is a compound having one ormore atoms of the compound replaced by an isotope of such atom. Forexample, homologs include compounds with deuterium in place of somehydrogen atoms of the compound such as compounds of the invention inwhich the methyl groups of the isopropoxy moiety of Formulas I-R and I-Sare fully or partially deuterated (e.g., (D₃C)₂C—O—). Isotopicsubstitutions which may be made in the formation of homologs of theinvention include non-radioactive (stable) atoms such as deuterium andcarbon 13, as well as radioactive (unstable) atoms such as tritium,carbon 14, iodine 123, iodine 125, etc.

A “solvate” is a similar composition except that a solvent other thatwater replaces the water. For example, methanol or ethanol can form an“alcoholate”, which can again be stoichiometic or non-stoichiometric. Asthe term is used herein a “solvate” refers to a solid form, i.e., acompound in solution in a solvent, while it may be solvated, is not asolvate as the term is used herein.

A “prodrug” as is well known in the art is a substance that can beadministered to a patient where the substance is converted in vivo bythe action of biochemicals within the patient's body, such as enzymes,to the active pharmaceutical ingredient. Examples of prodrugs includeesters of carboxylic acid groups, which can be hydrolyzed by endogenousesterases as are found in the bloodstream of humans and other mammals.

Any compound which can be converted in vivo to the active drug bychemical or biochemical transformations functions as a prodrug. Prodrugsof claimed compounds are covered under this invention.

Some examples of prodrugs within the scope of this invention include:

-   -   i. If the compound contains a hydroxyl group, the hydroxyl group        may be modified to form an ester, carbonate, or carbamate.        Examples include acetate, pivalate, methyl and ethyl carbonates,        and dimethylcarbamate. The ester may also be derived from amino        acids such as glycine, serine, or lysine.    -   ii. If the compound contains an amine group, the amine group may        be modified to form an amide. Examples include acetamide or        derivatization with amino acids such as glycine, serine, or        lysine.

Certain compounds of the invention and their salts may exist in morethan one crystal form and the present invention includes each crystalform and mixtures thereof. In addition, the compounds of the presentinvention can exist in unsolvated as well as solvated forms withpharmaceutically acceptable solvents such as water to form hydrates oradducts with alcohols such as C₁₋₄-alkanols, and the like. Furthermore,compounds of this invention can be isolated in association with solventmolecules by crystallization from evaporation of an appropriate solvent.Such solvents include but are not limited to toluene, tetrahydrofuran,dioxane, dimethylformamide, acetonitrile, acetates such as methylacetate, ethyl acetate, butyl acetate, isobutyl acetate, propyl- andisopropyl acetate, ethers such as diethyl ether and ethyl ether,alcohols such as methanol, ethanol, 1- or 2-butanol, 1- or 2-propanol,pentanol, and dimethylsulfoxide. In general, a depiction for thecompound by structure or name is considered to embrace the compound inany form (e.g., by itself, as a hydrate, solvate, or otherwise in amixture).

In addition, where features or aspects of the invention are described interms of Markush groups, those skilled in the art will recognize thatthe invention is also thereby described in terms of any individualmember or subgroup of members of the Markush group. For example, if X isdescribed as selected from the group consisting of bromine, chlorine,and iodine, claims for X being bromine and claims for X being bromineand chlorine are fully described. Moreover, where features or aspects ofthe invention are described in terms of Markush groups, those skilled inthe art will recognize that the invention is also thereby described interms of any combination of individual members or subgroups of membersof Markush groups. Thus, for example, if X is described as selected fromthe group consisting of bromine, chlorine, and iodine, and Y isdescribed as selected from the group consisting of methyl, ethyl, andpropyl, claims for X being bromine and Y being methyl are fullydescribed.

Compositions and Combination Treatments

The S1P₁ compounds, their pharmaceutically acceptable salts orhydrolyzable esters of the present invention may be combined with apharmaceutically acceptable carrier to provide pharmaceuticalcompositions useful for treating the biological conditions or disordersnoted herein in mammalian species, and more preferably, in humans. Theparticular carrier employed in these pharmaceutical compositions mayvary depending upon the type of administration desired (e.g.intravenous, oral, topical, suppository, or parenteral).

In preparing the compositions in oral liquid dosage forms (e.g.,suspensions, elixirs and solutions), typical pharmaceutical media, suchas water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents and the like can be employed. Similarly, when preparingoral solid dosage forms (e.g., powders, tablets and capsules), carrierssuch as starches, sugars, diluents, granulating agents, lubricants,binders, disintegrating agents and the like can be employed.

Another aspect of an embodiment of the invention provides compositionsof the compounds of the invention, alone or in combination with anotherS1P₁ inhibitor or another type of therapeutic agent, or both. As setforth herein, compounds of the invention include stereoisomers,tautomers, solvates, hydrates, salts including pharmaceuticallyacceptable salts, and mixtures thereof. Compositions containing acompound of the invention can be prepared by conventional techniques,e.g. as described in Remington: The Science and Practice of Pharmacy,19th Ed., 1995, incorporated by reference herein. The compositions canappear in conventional forms, for example capsules, tablets, aerosols,solutions, suspensions or topical applications.

Typical compositions include a compound of the invention and apharmaceutically acceptable excipient which can be a carrier or adiluent. For example, the active compound will usually be mixed with acarrier, or diluted by a carrier, or enclosed within a carrier which canbe in the form of an ampoule, capsule, sachet, paper, or othercontainer. When the active compound is mixed with a carrier, or when thecarrier serves as a diluent, it can be solid, semi-solid, or liquidmaterial that acts as a vehicle, excipient, or medium for the activecompound. The active compound can be adsorbed on a granular solidcarrier, for example contained in a sachet. Some examples of suitablecarriers are water, salt solutions, alcohols, polyethylene glycols,polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin,lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar,cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin,acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid,fatty acids, fatty acid amines, fatty acid monoglycerides anddiglycerides, pentaerythritol fatty acid esters, polyoxyethylene,hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrieror diluent can include any sustained release material known in the art,such as glyceryl monostearate or glyceryl distearate, alone or mixedwith a wax.

The formulations can be mixed with auxiliary agents which do notdeleteriously react with the active compounds. Such additives caninclude wetting agents, emulsifying and suspending agents, salt forinfluencing osmotic pressure, buffers and/or coloring substancespreserving agents, sweetening agents or flavoring agents. Thecompositions can also be sterilized if desired.

The route of administration can be any route which effectivelytransports the active compound of the invention which inhibits theenzymatic activity of the focal adhesion kinase to the appropriate ordesired site of action, such as oral, nasal, pulmonary, buccal,subdermal, intradermal, transdermal or parenteral, e.g., rectal, depot,subcutaneous, intravenous, intraurethral, intramuscular, intranasal,ophthalmic solution or an ointment, the oral route being preferred.

For parenteral administration, the carrier will typically comprisesterile water, although other ingredients that aid solubility or serveas preservatives can also be included. Furthermore, injectablesuspensions can also be prepared, in which case appropriate liquidcarriers, suspending agents and the like can be employed.

For topical administration, the compounds of the present invention canbe formulated using bland, moisturizing bases such as ointments orcreams.

If a solid carrier is used for oral administration, the preparation canbe tabletted, placed in a hard gelatin capsule in powder or pellet formor it can be in the form of a troche or lozenge. If a liquid carrier isused, the preparation can be in the form of a syrup, emulsion, softgelatin capsule or sterile injectable liquid such as an aqueous ornon-aqueous liquid suspension or solution.

Injectable dosage forms generally include aqueous suspensions or oilsuspensions which can be prepared using a suitable dispersant or wettingagent and a suspending agent Injectable forms can be in solution phaseor in the form of a suspension, which is prepared with a solvent ordiluent. Acceptable solvents or vehicles include sterilized water,Ringer's solution, or an isotonic aqueous saline solution.Alternatively, sterile oils can be employed as solvents or suspendingagents. Preferably, the oil or fatty acid is non-volatile, includingnatural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the formulation can also be a powder suitable forreconstitution with an appropriate solution as described above. Examplesof these include, but are not limited to, freeze dried, rotary dried orspray dried powders, amorphous powders, granules, precipitates, orparticulates. For injection, the formulations can optionally containstabilizers, pH modifiers, surfactants, bioavailability modifiers andcombinations of these. The compounds can be formulated for parenteraladministration by injection such as by bolus injection or continuousinfusion. A unit dosage form for injection can be in ampoules or inmulti-dose containers.

The formulations of the invention can be designed to provide quick,sustained, or delayed release of the active ingredient afteradministration to the patient by employing procedures well known in theart. Thus, the formulations can also be formulated for controlledrelease or for slow release.

Compositions contemplated by the present invention can include, forexample, micelles or liposomes, or some other encapsulated form, or canbe administered in an extended release form to provide a prolongedstorage and/or delivery effect. Therefore, the formulations can becompressed into pellets or cylinders and implanted intramuscularly orsubcutaneously as depot injections. Such implants can employ known inertmaterials such as silicones and biodegradable polymers, e.g.,polylactide-polyglycolide. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides).

For nasal administration, the preparation can contain a compound of theinvention which inhibits the enzymatic activity of the focal adhesionkinase, dissolved or suspended in a liquid carrier, preferably anaqueous carrier, for aerosol application. The carrier can containadditives such as solubilizing agents, e.g., propylene glycol,surfactants, absorption enhancers such as lecithin (phosphatidylcholine)or cyclodextrin, or preservatives such as parabens.

For parenteral application, particularly suitable are injectablesolutions or suspensions, preferably aqueous solutions with the activecompound dissolved in polyhydroxylated castor oil.

Dosage forms can be administered daily, or more than once a day, such astwice or thrice daily. Alternatively dosage forms can be administeredless frequently than daily, such as every other day, or weekly, if foundto be advisable by a prescribing physician.

An embodiment of the invention also encompasses prodrugs of a compoundof the invention which on administration undergo chemical conversion bymetabolic or other physiological processes before becoming activepharmacological substances. Conversion by metabolic or otherphysiological processes includes without limitation enzymatic (e.g,specific enzymatically catalyzed) and non-enzymatic (e.g., general orspecific acid or base induced) chemical transformation of the prodruginto the active pharmacological substance. In general, such prodrugswill be functional derivatives of a compound of the invention which arereadily convertible in vivo into a compound of the invention.Conventional procedures for the selection and preparation of suitableprodrug derivatives are described, for example, in Design of Prodrugs,ed. H. Bundgaard, Elsevier, 1985.

In another embodiment, there are provided methods of making acomposition of a compound described herein including formulating acompound of the invention with a pharmaceutically acceptable carrier ordiluent. In some embodiments, the pharmaceutically acceptable carrier ordiluent is suitable for oral administration. In some such embodiments,the methods can further include the step of formulating the compositioninto a tablet or capsule. In other embodiments, the pharmaceuticallyacceptable carrier or diluent is suitable for parenteral administration.In some such embodiments, the methods further include the step oflyophilizing the composition to form a lyophilized preparation.

The compounds of the invention can be used therapeutically incombination with i) one or more other S1P₁ inhibitors and/or ii) one ormore other types of protein kinase inhibitors and/or one or more othertypes of therapeutic agents which can be administered orally in the samedosage form, in a separate oral dosage form (e.g., sequentially ornon-sequentially) or by injection together or separately (e.g.,sequentially or non-sequentially).

Accordingly, in another embodiment the invention provides combinations,comprising:

-   -   a) a compound of the invention as described herein; and    -   b) one or more compounds comprising:        -   i) other compounds of the present invention,        -   ii) other medicaments adapted for treatment of a            malcondition for which activation of S1P₁ is medically            indicated, for example multiple sclerosis, transplant            rejection, or adult respiratory distress syndrome.

Combinations of the invention include mixtures of compounds from (a) and(b) in a single formulation and compounds from (a) and (b) as separateformulations. Some combinations of the invention can be packaged asseparate formulations in a kit. In some embodiments, two or morecompounds from (b) are formulated together while a compound of theinvention is formulated separately.

The dosages and formulations for the other agents to be employed, whereapplicable, will be as set out in the latest edition of the Physicians'Desk Reference, incorporated herein by reference.

Methods of Treatment

In certain embodiments, the present invention encompasses orallybioavailable compounds that specifically agonize SIP_(′) without binding(S1P₂, S1P₃ and S1P₄), or having significant specificity over (S1P₅),other EDG receptors. A selective S1P₁ agonist can be used to treatdiseases with an autoimmune, hyperactive immune-response, angiogenesisor inflammatory components, but would not be limited to such conditions.Selective S1P₁ agonists have advantages over current therapies byincreasing the therapeutic window because of reduced toxicity due toengagement of other EDG receptors.

In certain embodiments, the present invention encompasses compounds thatbind with high affinity and specificity to the S1P₁ receptor in anagonist manner. Upon ligation of the S1P₁ receptor with agonist,signaling proceeds through G_(αi), inhibiting the generation of cAMP byadenylate cyclase.

In certain embodiments, the present invention provides a method foractivating or agonizing (i.e., to have an agonic effect, to act as anagonist) a sphingosine-1-phosphate receptor subtype, such as S1P₁, witha compound of the invention. The method involves contacting the receptorwith a suitable concentration of an inventive compound to bring aboutactivation of the receptor. The contacting can take place in vitro, forexample in carrying out an assay to determine the S1P receptoractivation activity of an inventive compound undergoing experimentationrelated to a submission for regulatory approval.

In certain embodiments, the method for activating an S1P receptor, suchas S1P₁, can also be carried out in vivo, that is, within the livingbody of a mammal, such as a human patient or a test animal. Theinventive compound can be supplied to the living organism via one of theroutes as described above, e.g., orally, or can be provided locallywithin the body tissues, for example by injection of a tumor within theorganism. In the presence of the inventive compound, activation of thereceptor takes place, and the effect thereof can be studied.

An embodiment of the present invention provides a method of treatment ofa malcondition in a patient for which activation of an S1P receptor,such as S1P₁, is medically indicated, wherein the patient isadministered the inventive compound in a dosage, at a frequency, and fora duration to produce a beneficial effect on the patient. The inventivecompound can be administered by any suitable means, examples of whichare described above.

Preparation of Certain Embodiments

-   -   Reagents: (i) Zn(CN)₂, Pd(PPh₃)₄, NMP; (ii)        RuCl(p-cymene)[(R,R)-Ts-DPEN], HCO₂H-TEA complex; (iii)        NH₂OH*HCl, Na₂CO₃ or TEA, EtOH; (iv) HOBt, EDC, benzoic acid,        DMF.

The (S)-enantiomer was prepared in the same manner outlined in Scheme 1using RuCl(p-cymene)[(S,S)-Ts-DPEN] in step (ii). Racemic material canbe prepared in the same manner outlined in Scheme 1 using NaBH₄ in (ii).

-   -   Reagents: (i) DPPA, DBU, toluene; (ii) PG=protecting group e.g.        Boc: Pd/C, H₂, Boc₂O, TEA, MeOH; (iii) NH₂OH*HCl, NaHCO₃,        EtOH; (iv) HOBt, EDC, benzoic acid, DMF (v) deprotection e.g. 4M        HCl in dioxane; (vi) (a) R′-LG or R″-LG, where LG represents a        leaving group, K₂CO₃, CH₃CN; (b) R¹—CO₂H or R²—CO₂H, HOBt, EDC,        DMF or R¹—COCl or R²—COCl, TEA, DCM; (c) R¹—SO₂Cl or R³—SO₂Cl,        TEA, DCM (d) R²—CHO, HOAc, NaBH₄ or NaCNBH₃ or Na(OAc)₃BH,        MeOH; (e) R¹—OCOCl or R²—OCOCl, DIEA, DMF; (f) HN(R⁵R⁵), CDI,        TEA, DCM; (g) H₂NSO₂NH₂, Δ, dioxane; (h) dimethyloxirane, Δ,        EtOH; (vii) (a) If R′ or R″═H, then reactions (vi)(a-d) can be        performed; (b) If R′ or R″ contains an ester then (i) hydrolysis        NaOH, EtOH or (ii) reduction NaBH₄, MeOH can be performed; (c)        If R′ or R″ contains an acid then couplings HN(R⁵R⁵), HOBt, EDC,        DMF can be performed; (d) If R′ or R″ contains an appropriate        activated alkene then Michael additions HN(R⁵R⁵), DMF can be        performed.

The (R)-enantiomer was prepared in the same manner outlined in Scheme 2starting from(S)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile.

-   -   Reagents: (i) Sodium borohydride, ethanol, silica gel; (ii)        PG=protecting group e.g. TBDMS chloride, imidazole; (iii)        4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane),        PdCl₂(dppf).CH₂Cl₂, potassium acetate, dioxane.

-   -   Reagents: (i) Zn(CN)₂, Pd(PPh₃)₄, NMP; (ii) For racemic        material: Sodium borohydride, ethanol, silica gel; For        (R)-indanol: (S)-(−)-2-methyl-CBS-oxazaborolidine, BH₃-DMS,        toluene; For (S)-indanol: (R)-(+)-2-methyl-CBS-oxazaborolidine,        BH₃-DMS, toluene; (iii) NH₂OH*HCl, Na₂CO₃ or TEA, EtOH.

-   -   Reagents: (i) Oxalylchloride, DCM; (ii) Ethanolamine, Et₃N,        DCM; (iii) SOCl₂, DCM, KOH, MeOH (iv)N-Bromosuccinimide,        azoisobutyronitrile, DCM; (v) Protected (e.g. TBDMS)        4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ol,        K₂CO₃, Pd(PPh₃)₄, DME, H₂O; (vi) deprotection e.g. TBAF,        THF; (vii) SOCl₂, DCM; (viii) R′-NH₂ or R″—NH₂, DIPEA, DMA.

-   -   Reagents: (i) (R)-2-methylpropane-2-sulfinamide, Ti(OEt)₄,        toluene; (ii) NaBH₄, THF; (iii) 4N HCl in dioxane, MeOH; (iv)        Boc₂O, TEA, DCM; (v)        4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane),        PdCl₂(dppf).CH₂Cl₂, potassium acetate, dioxane; (vi)        5-(5-bromooxazol-2-yl)-2-isopropoxybenzonitrile, K₂CO₃,        Pd(PPh₃)₄, DME, H₂O; (vii) 4N HCl in dioxane; (viii) (a) R′-LG        or R″-LG, where LG represents a leaving group, K₂CO₃, CH₃CN; (b)        R¹—CO₂H or R²—CO₂H, HOBt, EDC, DMF or R¹—COCl or R²—COCl, TEA,        DCM; (c) R¹—SO₂Cl or R³—SO₂Cl, TEA, DCM (d) R²—CHO, HOAc, NaBH₄        or NaCNBH₃ or Na(OAc)₃BH, MeOH; (e) R¹—OCOCl or R²—OCOC1, DIEA,        DMF; (f) HN(R⁵R⁵), CDI, TEA, DCM; (g) H₂NSO₂NH₂, Δ, dioxane; (h)        dimethyloxirane, Δ, EtOH; (ix) (a) If R′ or R″═H, then reactions        (viii)(a-d) can be performed; (b) If R′ or R″ contains an ester        then (i) hydrolysis NaOH, EtOH or (ii) reduction NaBH₄, MeOH can        be performed; (c) If R′ or R″ contains an acid then couplings        HN(R⁵R⁵), HOBt, EDC, DMF can be performed; (d) If R′ or R″        contains an appropriate activated alkene then Michael additions        HN(R⁵R⁵), DMF can be performed.

The (S)-enantiomer can be prepared using(S)-2-methylpropane-2-sulfinamide in step (i).

-   -   Reagents: (i) HOBt, EDC, 2-(3,4-diethoxyphenyl)acetic acid,        DMF; (ii) SOCl₂, DCM; (iii) R′-NH₂, DIPEA, DMA.

-   -   Reagents: (i) Zn(CN)₂, Pd(PPh₃)₄, NMP; (ii)        (R)-2-methylpropane-2-sulfinamide, Ti(OEt)₄, toluene; (iii)        NaBH₄, THF; (iv) 4M HCl in dioxane, MeOH; (v) PG=protecting        group e.g. Boc₂O, TEA, DCM; (vi) NH₂OH*HCl, TEA, EtOH; (vii)        R′-halide, NaH, DMF.

-   -   Reagents: (i) (a) HOBt, EDC, 2-(3,4-diethoxyphenyl)acetic acid,        DMF (b) deprotection e.g. 4N HCl in dioxane; (ii) (a) R′-LG,        where LG represents a leaving group, K₂CO₃, CH₃CN; (b) if R′        contains an ester then (a) followed by NaOH, EtOH; (c) R′-CO₂H,        HOBt, EDC, DMF or R′-COCl, TEA, DCM; (d) R′-SO₂Cl, TEA, DCM (e)        R′-CHO, HOAc, NaBH₄ or NaCNBH₃ or Na(OAc)₃BH, MeOH.

The (S)-enantiomer can be prepared using protected(R)-1-amino-N-hydroxy-2,3-dihydro-1H-indene-4-carboximidamide in step(i).

-   -   Reagents: (i) HOBt, EDC,        4-phenyl-5-(trifluoromethyl)thiophene-2-carboxylic acid,        DMF; (ii) 2N HCL in ether, DCM.

-   -   Reagents: (i) PG=protecting group e.g. Boc₂O, DMAP, ACN; (ii)        NH₂OH*HCl, Na₂CO₃, EtOH; (iii) HOBt, EDC, benzoic acid,        DMF; (iv) deprotection e.g. 4N HCl in dioxane.

-   -   Reagents: (i) NH₂OH*HCl, Na₂CO₃, EtOH; (ii) HOBt, EDC, benzoic        acid, DMF.

-   -   Reagents: (i) NH₂OH*HCl, Na₂CO₃, EtOH; (ii) HOBt, EDC,        3-cyano-4-isopropoxybenzoic acid, DMF.

-   -   Reagents: (i) PG=protecting group e.g.        tert-butylchlorodimethylsilane, TEA, DCM; (ii) Zn(CN)₂,        Pd(PPh₃)₄, NMP; (iii) NH₂OH*HCl, Na₂CO₃, EtOH; (iv) HOBt, EDC,        benzoic acid, DMF.

Examples General Methods

¹H NMR (400 MHz) and ¹³C NMR (100 MHz) were obtained in solution ofdeuteriochloroform (CDCl₃), deuteriomethanol (CD₃OD) or dimethylsulfoxide—D₆ (DMSO). NMR spectra were processed using Mestrec 5.3.0 and6.0.1. ¹³C NMR peaks that are bracketed are two rotomers of the samecarbon. Mass spectra (LCMS) were obtained using an Agilent 1100/6110HPLC system equipped with a Thompson ODS-A, 100A, 5μ (50×4.6 mm) columnusing water with 0.1% formic acid as the mobile phase A, andacetonitrile with 0.1% formic acid as the mobile phase B. The gradientwas 20-100% with mobile phase B over 2.5 min then held at 100% for 2.5mins. The flow rate was 1 mL/min. Unless otherwise indicated, the LCMSdata provided uses this method. For more hydrophobic compounds, thefollowing gradient was used, denoted as Method 1: 40-95% over 0.5 min,hold at 95% for 8.5 min, with a flow rate of 1 mL/min. Final compoundswere checked for purity using Method 2: 5% for 1 min, 5-95% over 9 min,then hold at 95% for 5 min, with a flow rate of 1 mL/min. Enantiomericexcess was determined by integration of peaks that were separated on aChiralpak AD-H, 250×4.6 mm column at a flow rate of 1 mL/min and anisocratic mobile phase. Unless otherwise indicated, the chiral dataprovided uses this method. Alternatively, chiral separations wereperformed under the following conditions, denoted as Chiral Method 1:Chiralpak AY-H, 250×4.6 mm column at a flow rate of 1 mL/min and anisocratic mobile phase. Chiral Method 2: Chiralcel OZ-3, 150×4.6 mmcolumn at a flow rate of 0.75 ml/min and an isocratic mobile phase. Thepyridine, dichloromethane (DCM), tetrahydrofuran (THF), and toluene usedin the procedures were from Aldrich Sure-Seal bottles kept undernitrogen (N₂). All reactions were stirred magnetically and temperaturesare external reaction temperatures. Chromatographies were carried outusing a Combiflash Rf flash purification system (Teledyne Isco) equippedwith Redisep (Teledyne Isco) silica gel (SiO₂) columns. Preparative HPLCpurifications were done on Varian ProStar/PrepStar system using watercontaining 0.05% trifluoroacetic acid as mobile phase A, andacetonitrile with 0.05% trifluoroacetic acid as mobile phase B. Thegradient was 10-80% with mobile phase B over 12 min, hold at 80% for 2min, and then return to 10% over 2 min with flow rate of 22 mL/min.Other methods similar to this may have been employed. Fractions werecollected using a Varian Prostar fraction collector and were evaporatedusing a Savant SpeedVac Plus vacuum pump. Compounds with salt-ablecenters were presumed to be the trifluoroacetic acid (TFA) salt.Microwave heating was performed using a Biotage Initiator microwavereactor equipped with Biotage microwave vessels. The followingabbreviations are used: ethyl acetate (EA), triethylamine (TEA), diethylamine (DEA), diispropyl ethyl amine (DIEA), hydroxybenzotriazole (HOBt),1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC),isopropanol (IPA), dimethylformamide (DMF), dimethyl acetamide (DMA).Norit is activated charcoal.

Experimental Procedures5-oxo-5,6,7,8-tetrahydronaphthalene-1-carbonitrile (INT-1)

To a stirred solution of 5-bromo-3,4-dihydronaphthalen-1(2H)-one (9.95g, 44.2 mmol) in NMP (50 mL) was added Zn(CN)₂ (10.38 g, 88.4 mmol). Themixture was degassed twice by bubbling N₂ through the solution for 30min then evacuated. Pd(Ph₃)₄ (0.5 g, 0.44 mmol) was added and themixture was heated to 110° C. under N₂. After 5 h, the mixture wascooled to room temperature and poured onto ice (600 mL), using water(300 mL) to complete the transfer. After the ice had melted, thesolution was filtered and the resulting solid was collected, suspendedin DCM, and filtered again. The solid was collected, washed with water,and purified by column chromatography (EA/hex) to provide 6.9 g (91%) of5-oxo-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-1 as a whitesolid. LCMS-ESI (m/z) calculated for C₁₁H₉NO: 171.2. found 172.1 [M+H]⁺,t_(R)=2.95 min. ¹H NMR (400 MHz, CDCl₃) δ 8.26 (dd, J=7.9, 1.4 Hz, 1H),7.82 (dd, J=7.6, 1.4 Hz, 1H), 7.44 (t, J=7.8 Hz, 1H), 3.20 (t, J=6.1 Hz,2H), 2.72 (dd, J=7.2, 6.1 Hz, 2H), 2.30-2.17 (m, 2H). ¹³C NMR (101 MHz,CDCl₃) δ 196.22, 147.39, 137.18, 133.39, 131.59, 127.19, 116.93, 112.94,38.48, 28.05, 22.28.

(R)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile (INT-2)

To a stirred solution of5-oxo-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-1 (3.0 g, 17.5mmol) in 5:1 HCO₂:NEt₃ (24 mL) was added RuCl(p-cymene)[(R,R)-Ts-DPEN](0.13 g, 0.26 mmol). The mixture was stirred at 30° C. for 15 h thenpartitioned between EA and H₂O. The combined organic layers were driedover Na₂SO₄ and chromatographed (EA/hex) to provide 2.99 g (99%) of(R)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-2 as awhite solid. LCMS-ESI (m/z) calculated for C₁₁H₁₁NO: 173.2. found 174.1[M+H]⁺, 156.1 [M-NH₄]⁺, t_(R)=2.60 min. ¹H NMR (400 MHz, CDCl₃) δ 7.71(d, J=7.8 Hz, 1H), 7.54 (dt, J=8.7, 4.4 Hz, 1H), 7.34-7.26 (m, 1H),4.85-4.71 (m, 2H), 3.48 (s, 1H), 3.13-2.96 (m, 1H), 2.90 (ddd, J=17.7,7.8, 5.6 Hz, 1H), 2.15-1.95 (m, 2H), 1.97-1.76 (m, 2H). Chiral HPLC:(R)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile was elutedwith 5% IPA/hexane: 99.1% ee, t_(R)=15.3 min.

(S)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-3 wasprepared in an analogous fashion using INT-1 andRuCl(p-cymene)[(S,S)-Ts-DPEN]. Chiral HPLC: 99.4% ee, t_(R) for the(S)-enantiomer=17.99 min.

General Procedure 1. Preparation of Amide Oximes

To (R)- or (S)-cyanides (1 eq) in EtOH (0.56 M) was added hydroxylaminehydrochloride (3 eq) and either NaHCO₃ or TEA (3 eq) and the reactionmixture heated at 85° C. for 1-2 h. The organic soluble amide oximeswere isolated by removal of the solvent and partitioning between waterand DCM. The water soluble amide oximes were chromatographed or useddirectly in the cyclization. Pure amide oximes can be obtained byrecrystallization from alcoholic solvents.

(R)—N,5-dihydroxy-5,6,7,8-tetrahydronaphthalene-1-carboximidamide(INT-4)

Prepared using General Procedure 1. To a stirring solution of(R)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-2 (79.1mg, 0.46 mmol) in EtOH (2 mL) was added hydroxylamine hydrochloride(34.9 mg, 0.50 mmol) and sodium bicarbonate (42.2 mg, 0.50 mmol). Themixture was heated at 70° C. for 18 h. The product was purified bychromatography (MeOH/DCM) to provide 27.3 mg (29%)(R)—N,5-dihydroxy-5,6,7,8-tetrahydronaphthalene-1-carboximidamide INT-4as a white solid. LCMS-ESI (m/z) calculated for C₁₁H₁₁NO: 173.2. found174.1 [M+H]⁺, 156.1 [M−NH₄]⁺, t_(R)=2.60min.(S)—N,5-dihydroxy-5,6,7,8-tetrahydronaphthalene-1-carboximidamideINT-5 was prepared in an analogous fashion from(S)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-3.

General Procedure 2. Cyclization to Oxadiazole Amines

A solution of the appropriate acid (1 eq), HOBt (1.3 eq), and EDC (1.3eq) in DMF (0.08 M in acid) was stirred at room temperature under anatmosphere of N₂. After the complete formation of the HOBt-acid complex(1-3 h), the (R)- or (S)-amide oxime (1.1 eq) was added to the mixture.After complete formation of the coupled intermediate (ca. 0.5-2 h), themixture was heated to 75-95° C. until the cyclization was complete (8-12h). The reaction mixture was diluted with saturated NaHCO₃ and extractedwith EA. The combined organic extracts were dried, concentrated, andcould be purified by chromatography (EA/hexanes), preparative HPLC orrecrystallization.

(R)-5-(3-(5-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile(Compound 1)

Prepared using General Procedure 2. To a stirring solution of3-cyano-4-isopropoxybenzoic acid (16.7 mg, 0.08 mmol) in DMF (1 mL) wereadded HOBt (14.3 mg, 0.11 mmol) and EDCI (20.3 mg, 0.11 mmol). Afterstirring for 30 min,(R)—N,5-dihydroxy-5,6,7,8-tetrahydronaphthalene-1-carboximidamide INT-4(27.3 mg, 0.09 mmol) was added as a solution in DMF (1.5 mL). Afterstirring at room temperature for an additional 60 min, the mixture washeated to 90° C. for 15 h. The mixture was diluted with EA and washedwith NaHCO₃. The combined organic layers were dried, concentrated,chromatographed (EA/hexanes) to provide 12.72 mg (42.4%)(R)-5-(3-(5-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile1 as a white solid. LCMS-ESI (m/z) calculated for C₂₂H₂₁N₃O₃: 375.4.found 376.1 [M+H]⁺, t_(R)=3.73 min. ¹H NMR (400 MHz, CDCl₃) δ 8.42 (d,J=2.2 Hz, 1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 7.97 (dd, J=7.7, 1.3 Hz,1H), 7.66 (d, J=7.2 Hz, 1H), 7.38 (t, J=7.7 Hz, 1H), 7.12 (d, J=9.0 Hz,1H), 4.91-4.83 (m, 1H), 4.79 (dq, J=12.0, 6.0 Hz, 1H), 3.20 (dt, J=17.8,5.4 Hz, 1H), 3.01 (dt, J=13.3, 6.4 Hz, 1H), 2.13-1.81 (m, 4H), 1.79 (d,J=7.2 Hz, 1H), 1.47 (d, J=5.6 Hz, 6H). ¹³C NMR (101 MHz, CDCl₃) δ172.70, 169.48, 162.75, 140.10, 137.4, 134.13, 133.88, 131.68, 129.96,126.18, 125.97, 116.82, 115.26, 113.54, 103.95, 72.73, 68.47, 31.62,28.50, 21.73, 18.57. Chiral HPLC:(R)-5-(3-(5-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilewas eluted with 10% IPA/hexane: 99.4% ee, t_(R)=40.85 min.

(S)-5-(3-(5-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile2 was prepared in an analogous fashion from(S)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-5. ChiralHPLC: 99.1% ee, t_(R) for the (S)-enantiomer=38.19 min.

(S)-5-azido-5,6,7,8-tetrahydronaphthalene-1-carbonitrile (INT-6)

A stirring solution of(R)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-2 (3.00g, 17.32 mmol) in toluene (16 mL) under N₂, was cooled to 0° C. DPPA(9.53 g, 34.64 mmol) was added, followed by dropwise addition of DBU(3.16 mL, 20.78 mmol) over 20 min. The mixture was stirred at 0° C. for4 h then slowly warmed to room temperature over 2 h and thenconcentrated. The resulting crude mixture was diluted with EA and washedwith NaHCO₃. The combined organic layers were washed with brine, driedover Na₂SO₄, and chromatographed (EA/hexane) to provide 2.49 g (72.6%)of (S)-5-azido-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-6 as awhite solid. LCMS-ESI (m/z) calculated for C₁₁H₁₀N₄: 198.2. found 156.1[M−N₃]⁺, t_(R)=3.65 min. ¹H NMR (400 MHz, CDCl₃) δ 7.60 (dd, J=7.6, 1.2Hz, 1H), 7.56 (dd, J=7.8, 0.6 Hz, 1H), 7.33 (t, J=7.7 Hz, 1H), 4.59 (t,J=4.4 Hz, 1H), 3.08 (dt, J=18.0, 5.1 Hz, 1H), 2.99-2.83 (m, 1H),2.15-1.96 (m, 3H), 2.00-1.81 (m, 1H). ¹³C NMR (101 MHz, DMSO) δ 141.05,135.58, 133.47, 132.66, 126.58, 117.43, 113.21, 58.74, 28.28, 27.54,18.27.

(R)-5-azido-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-7 wasprepared in an analogous fashion from(S)-5-hydroxy-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-3.

(S)-tert-butyl (5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate(INT-8)

To a stirring solution of(S)-5-azido-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-6 (3.1 g,15.63 mmol) in MeOH (50 mL) were added 10% Pd/C (500 mg), (Boc)₂O (6.83g, 31.27 mmol) and Et₃N (3.16 g, 31.27 mmol). The reaction mixture waspurged and flushed with H₂ (3×) and stirred under H₂. After 3 h themixture was filtered through celite, rinsing with MeOH. The MeOHfiltrate was concentrated, dissolved in EA and washed with NaHCO₃ andbrine. The organic layer was dried (MgSO₄), concentrated andchromatographed (EA/hexanes). The resulting material was crystallizedfrom hexanes to provide 3.45 g (81%) of (S)-tert-butyl(5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate INT-8 as a whitesolid. LCMS-ESI (m/z) calculated for C₁₆H₂₀N₂O₂: 272.34. found 156.1[M-NHBoc]⁺, t_(R)=3.77 min. ¹H NMR (400 MHz, CDCl₃) δ 7.60 (d, J=7.8 Hz,1H), 7.53 (d, J=7.5 Hz, 1H), 7.31-7.20 (m, 1H), 4.86 (s, 1H), 4.75 (d,J=8.9 Hz, 1H), 3.06-2.85 (m, 2H), 2.07 (dt, J=11.3, 5.1 Hz, 1H), 1.91(s, 2H), 1.86-1.71 (m, 1H), 1.48 (s, 9H). ¹³C NMR (101 MHz, CDCl₃) δ155.38, 140.70, 139.12, 133.03, 131.63, 126.41, 117.62, 112.42, 79.62,48.25, 29.75, 28.28, 27.77, 19.36. Chiral HPLC: (S)-tert-butyl(5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate was eluted with2.5% EtOH/hexanes: 92.4% ee, t_(R)=14.22 min.

(R)-tert-butyl (5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-9 was prepared in an analogous fashion from(R)-5-azido-5,6,7,8-tetrahydronaphthalene-1-carbonitrile INT-7. ChiralHPLC: 99.6% ee, t_(R) for the (R)-enantiomer=11.60 min.

(S)-tert-butyl(5-(N-hydroxycarbamimidoyl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate(INT-10)

Prepared using General Procedure 1. To a stirring solution of(S)-tert-butyl (5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-8 (3.10 g, 11.38 mmol) in EtOH (25 mL) was added hydroxylaminehydrochloride (2.77 g, 39.84 mmol) and NEt₃ (3.17 mL, 22.77 mmol). Afterheating at 85° C. for 15 h, the mixture was concentrated, redissolved inDCM, and washed with NaHCO₃. The combined organic layers were dried,concentrated and chromatographed (MeOH/DCM) to provide 3.56 g crude(S)-tert-butyl(5-(N-hydroxycarbamimidoyl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-10 (58% product by UV area), which was used in the next reactionwithout further purification. LCMS-ESI (m/z) calculated for C₁₆H₂₃N₃O₃:305.37. found 306.2 [M+H]⁺, t_(R)=2.00 min.

(R)-tert-butyl(5-(N-hydroxycarbamimidoyl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-11 was prepared in an analogous fashion from (R)-tert-butyl(5-cyano-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate INT-9.

(S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate(INT-12)

Prepared using General Procedure 2. To a stirring solution of3-cyano-4-isopropoxybenzoic acid (556 mg, 2.72 mmol) in DMF (10 mL) wasadded HOBt (476.6 mg, 3.53 mmol) and EDCI (677.9 mg, 3.53 mmol). Afterstirring for 30 min, (S)-tert-butyl(5-(N-hydroxycarbamimidoyl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-10 (3.56 g crude, approximately 2.99 mmol) was added. After stirringat room temperature for an additional 90 min, the mixture was heated to90° C. for 15 h. The mixture was diluted with EA and washed with NaHCO₃.The combined organic layers were dried, concentrated, andchromatographed (EA/hexanes) to provide 1.89 g (46%) (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-12 as a white solid. LCMS-ESI (m/z) calculated for C₂₇H₃₀N₄O₄:474.6; no M/Z observed, t_(R)=4.23 min. ¹H NMR (400 MHz, CDCl₃) δ 8.42(d, J=2.1 Hz, 1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 7.92 (dd, J=7.7, 1.1Hz, 1H), 7.56 (d, J=7.6 Hz, 1H), 7.32 (dd, J=20.3, 12.6 Hz, 1H), 7.12(d, J=9.0 Hz, 1H), 4.94 (d, J=6.0 Hz, 1H), 4.88-4.72 (m, 1H), 3.23-3.08(m, 1H), 3.07-2.94 (m, 1H), 2.06 (d, J=12.6 Hz, 1H), 1.97-1.78 (m, 3H),1.53-1.43 (m, 15H). ¹³C NMR (101 MHz, CDCl₃) δ 172.64, 169.40, 162.69,155.43, 138.65, 137.55, 134.04, 133.83, 131.68, 129.55, 126.08, 125.97,116.74, 115.20, 113.53, 103.87, 79.44, 72.69, 48.97, 29.73, 28.40,21.68, 19.71, 14.14.

(R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-13 was prepared in an analogous fashion from (R)-tert-butyl(5-(N-hydroxycarbamimidoyl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-11.

(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile(Compound 4)

To a stirring solution of (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamateINT-12 (0.90 g, 1.9 mmol) in dioxanes (10 mL) was added 4N HCl/dioxanes(2.5 mL). After stirring at 60° C. for 5.5 h, the mixture wasconcentrated to provide 0.8 g (100%) of the HCl salt of(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile4 as a white solid. An analytically pure sample was purified bypreparative HPLC and the free amine was prepared by partitioning betweenNaHCO₃ and EA. LCMS-ESI (m/z) calculated for C₂₂H₂₂N₄O₂: 374.4. found358.1 [M−NH₂]⁺, t_(R)=2.45 min. ¹H NMR (400 MHz, DMSO) δ 8.63 (s, 2H),8.50 (d, J=2.2 Hz, 1H), 8.39 (dd, J=9.0, 2.3 Hz, 1H), 7.95 (dd, J=7.7,1.0 Hz, 1H), 7.85 (d, J=7.5 Hz, 1H), 7.56 (d, J=9.2 Hz, 1H), 7.49 (t,J=7.7 Hz, 1H), 4.98 (hept, J=6.0 Hz, 1H), 4.55 (t, J=5.3 Hz, 1H), 3.11(dt, J=17.9, 5.5 Hz, 1H), 2.94 (dt, J=13.9, 6.1 Hz, 1H), 2.18-1.88 (m,3H), 1.88-1.71 (m, 1H), 1.38 (d, J=6.0 Hz, 6H). ¹³C NMR (101 MHz, DMSO)δ 172.91, 168.60, 162.52, 137.73, 134.60, 134.09, 133.80, 132.08,130.30, 126.29, 126.06, 115.92, 115.24, 114.93, 102.49, 72.54, 66.34,48.02, 27.57, 26.54, 21.47, 17.68. Chiral HPLC:(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilewas eluted with 8% EtOH/hexanes, with 0.3% DEA (Chiral Method 1): 94.2%ee, t_(R)=42.7 min.

(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile3 was prepared in an analogous fashion from (R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)carbamate INT-13. Chiral HPLC (Chiral Method 1): 99.9% ee, t_(R) for the(R)-enantiomer=39.72 min.

General Procedure 3. Preparation of Tetrahydronaphthalene Ureas

To a stirring solution of CDI (1.2 eq) in DCM (0.16M) were added eitherthe solution of (R)- or (S)-tetrahydronapthalene amine (1 eq) and Et₃N(3 eq) in DCM (0.01M). After stirring for 15 h, this solution was addedto a second solution containing the appropriate amine (3 eq) and Et₃N (3eq) in DCM (0.4M), at room temperature. The resulting mixture wasstirred at room temperature for 4 h until all of starting material wasconsumed. The solvent was evaporated and the pure product isolated afterpreparative HPLC.

Compounds 5-20 were prepared using General Procedure 3.

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-3-hydroxyazetidine-1-carboxamide(Compound 8)

Prepared using General Procedure 3: LCMS-ESI (m/z) calculated forC₂₆H₂₇N₅O₄: 473.5. found 474.2 [M+H]⁺, t_(R)=3.21 min. ¹H NMR (400 MHz,CDCl₃) δ 8.41 (d, J=2.2 Hz, 1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 7.91 (dd,J=7.7, 1.2 Hz, 1H), 7.53 (t, J=6.1 Hz, 1H), 7.32 (dd, J=20.1, 12.4 Hz,1H), 7.12 (d, J=9.0 Hz, 1H), 5.13 (d, J=8.0 Hz, 1H), 4.79 (dt, J=12.2,6.1 Hz, 1H), 4.68 (tt, J=6.7, 4.4 Hz, 1H), 4.35 (d, J=8.7 Hz, 1H),4.25-4.14 (m, 2H), 3.85 (dd, J=8.8, 4.1 Hz, 2H), 3.14 (t, J=12.1 Hz,1H), 3.10-2.92 (m, 1H), 2.16-1.96 (m, 1H), 1.99-1.64 (m, 4H), 1.48 (d,J=6.1 Hz, 6H). Chiral HPLC:(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-3-hydroxyazetidine-1-carboxamidewas eluted with 15% water/MeOH, (Chiral Method 2): 91.4% ee, t_(R)=15.52min.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-3-hydroxyazetidine-1-carboxamide7 was prepared in an analogous fashion. Chiral HPLC: 99.94% ee,t_(R)=17.17 min (Chiral Method 2).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)pyrrolidine-1-carboxamide(Compound 12)

Prepared using General Procedure 3: LCMS-ESI (m/z) calculated forC₂₇H₂₉N₅O₃: 471.55. found 472.2 [M+H]⁺, t_(R)=3.78 min. ¹H NMR (400 MHz,CDCl₃) δ 8.41 (d, J=2.1 Hz, 1H), 8.32 (dd, J=8.9, 2.2 Hz, 1H), 7.90 (dd,J=7.6, 1.1 Hz, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.33 (t, J=7.7 Hz, 1H), 7.12(d, J=9.0 Hz, 1H), 5.25-5.12 (m, 1H), 4.79 (hept, J=6.0 Hz, 1H), 4.45(t, J=21.8 Hz, 1H), 3.36 (t, J=6.4 Hz, 4H), 3.23-3.09 (m, 1H), 3.02 (dt,J=18.0, 6.0 Hz, 1H), 2.16-1.99 (m, 1H), 2.01-1.79 (m, 7H), 1.47 (d,J=6.1 Hz, 6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)morpholine-4-carboxamide(Compound 15)

Prepared using General Procedure 3: LCMS-ESI (m/z) calculated forC₂₇H₂₉N₅O₄: 487.5. found 488.2 [M+H]⁺, t_(R)=3.58 min. ¹H NMR (400 MHz,CDCl₃) δ 8.40 (d, J=2.2 Hz, 1H), 8.32 (dd, J=8.9, 2.2 Hz, 1H), 7.91 (dd,J=7.7, 1.1 Hz, 1H), 7.55 (t, J=10.0 Hz, 1H), 7.33 (t, J=7.7 Hz, 1H),7.12 (d, J=9.0 Hz, 1H), 5.19 (dd, J=12.9, 5.3 Hz, 1H), 4.86-4.75 (m,1H), 4.72 (d, J=8.1 Hz, 1H), 3.76-3.63 (m, 4H), 3.36 (dd, J=11.6, 7.1Hz, 4H), 3.16 (dt, J=16.7, 5.4 Hz, 1H), 3.02 (dt, J=12.6, 5.9 Hz, 1H),2.14-1.98 (m, 2H), 1.94-1.81 (m, 2H), 1.47 (d, J=6.1 Hz, 6H).

(R)—N—((R)-5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-3-(dimethylamino)pyrrolidine-1-carboxamide(Compound 20)

Prepared using General Procedure 3: To a stirring solution of CDI ((9.5mg, 0.06 mmol) in DCM (1 mL) were added dropwise a solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrileHCl salt 3 (20 mg, 0.05 mmol) and Et₃N (20.3 μL, 0.15 mmol) in DCM (1mL). After stirring for 15 h at room temperature, this solution wasadded dropwise to another solution containing(R)-3-dimethylaminopyrrolidine (18.6 mg, 0.15 mmol)) in DCM (1 mL) atroom temperature. The reaction was stirred at room temperature for 6.5h. The solvent was evaporated and the pure product was isolated bypreparative HPLC to afford 17.5 mg (70%) of(R)—N—((R)-5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-3-(dimethylamino)pyrrolidine-1-carboxamide 20. LCMS-ESI (m/z) calculated for C₂₉H₃₄N₆O₃:514.6. found 515.3 [M+H]⁺, t_(R)=2.56 min. ¹H NMR (400 MHz, CDCl₃) δ8.39 (d, J=2.2 Hz, 1H), 8.31 (dt, J=8.7, 4.3 Hz, 1H), 7.91 (d, J=7.6 Hz,1H), 7.52 (d, J=7.6 Hz, 1H), 7.34 (t, J=7.7 Hz, 1H), 7.12 (d, J=9.2 Hz,1H), 5.13 (d, J=6.9 Hz, 1H), 4.86-4.73 (m, 1H), 4.64 (d, J=8.2 Hz, 1H),3.91 (dd, J=10.4, 7.3 Hz, 1H), 3.80-3.56 (m, 3H), 3.41 (dd, J=17.5, 8.3Hz, 1H), 3.15 (d, J=18.0 Hz, 1H), 3.10-2.93 (m, 1H), 2.85 (s, 6H), 2.46(m, 2H), 2.05 (dd, J=9.2, 4.8 Hz, 1H), 1.88 (m, 3H), 1.45 (dd, J=13.9,6.1 Hz, 6H).

General Procedure 4. Preparation of Tetrahydronapthalene Amides via AcidChlorides

To a stirring solution of (R)- or (S)-tetrahydronapthalene amine HCl (1eq) in DCM were added an acid chloride (2 eq) and NEt₃ (2 eq). Thereaction was stirred at room temperature for 1 h. The solvent wasevaporated and mixture was purified by preparative HPLC.

Compounds 21-25 were prepared using General Procedure 4.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)acetamide(Compound 21)

Prepared using General Procedure 4: To a stirring solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrileHCl 3 (20 mg, 0.05 mmol) in DCM (0.5 mL) were added acetyl chloride (7μL, 0.10 mmol) and NEt₃ (14 μL, 0.10 mmol). After stirring for 1 h, thesolvent was evaporated and the residue was purified by preparative HPLCto provide 11.3 mg (56%) of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)acetamide21. LCMS-ESI (m/z) calculated for C₂₄H₂₄N₄O₃: 416.5. found 417.2 [M+H]⁺,t_(R)=3.56 min. ¹H NMR (400 MHz, CDCl₃) δ 8.40 (d, J=2.2 Hz, 1H), 8.32(dd, J=8.9, 2.2 Hz, 1H), 7.92 (dd, J=7.7, 1.2 Hz, 1H), 7.48 (d, J=7.6Hz, 1H), 7.33 (t, J=7.7 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H), 5.83 (d, J=8.6Hz, 1H), 5.36-5.21 (m, 1H), 4.79 (hept, J=6.0 Hz, 1H), 3.16 (dt, J=17.9,6.0 Hz, 1H), 3.03 (dt, J=18.2, 6.3 Hz, 1H), 2.10-1.99 (m, 4H), 1.97-1.79(m, 3H), 1.47 (d, J=6.1 Hz, 6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)acetamide22 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrileHCl 4.

General Procedure 5. Preparation of Tetrahydronapthalene Sulfamides

To a solution of (R)- or (S)-tetrahydronapthalene amine HCl (1 eq) indioxane were added sulfamide (5 eq) and DIEA (3 eq). The reaction wasstirred at 110° C. for 18 h. The solvent was evaporated and mixture waspurified by preparative HPLC.

Compounds 26 and 27 were prepared using General Procedure 5.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamide(Compound 26)

Prepared using General Procedure 5: To a stirring solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrileHCl 3 (50 mg, 0.12 mmol) in dioxane (3 mL) were added sulfamide (58 mg,0.61 mmol) and DIEA (47.2 μL, 0.37 mmol) and the mixture was heated to110° C. for 14 h. The solvent was evaporated and the residue waspurified by preparative HPLC to provide 22.8 mg (42%) of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamide26. LCMS-ESI (m/z) calculated for C₂₂H₂₃N₅O₄S: 453.5. found 454.1[M+H]⁺, t_(R)=3.47 min. ¹H NMR (400 MHz, CDCl₃) δ 8.41 (d, J=2.2 Hz,1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 7.97 (dd, J=7.7, 1.2 Hz, 1H), 7.72(d, J=7.7 Hz, 1H), 7.38 (t, J=7.8 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H), 4.78(ddd, J=13.4, 11.7, 5.7 Hz, 2H), 4.59 (d, J=19.8 Hz, 2H), 4.55 (d, J=8.2Hz, 1H), 3.19 (dt, J=18.0, 5.6 Hz, 1H), 3.02 (dt, J=18.2, 7.2 Hz, 1H),2.23-2.03 (m, 2H), 1.92 (dt, J=12.4, 6.3 Hz, 2H), 1.48 (d, J=6.1 Hz,6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamide27 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrileHC14.

General Procedure 6. Preparation of Tetrahydronaphthalene Sulfonamidesvia Sulfonyl Chlorides

To a solution of (R)- or (S)-tetrahydronapthalene amine HCl (1 eq) inDCM (0.05M) was added TEA (2 eq) and the appropriate sulfonyl chloride(1-2 eq.) at room temperature. The reaction mixture was stirred at roomtemperature for 18 h. The solvent was evaporated and the productisolated after preparative HPLC purification.

Compounds 28-33 were prepared using General Procedure 6.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)methanesulfonamide(Compound 28)

Prepared using General Procedure 6: To a stirring solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 3 (20 mg, 0.05 mmol) in DCM (1 mL) at 0° C. was added TEA(20 μL, 0.15 mmol) and methanesulfonyl chloride (4.5 μL, 0.06 mmol). Themixture was allowed to warm to room temperature over 2 h. The solventwas evaporated and crude mixture was purified by preparative HPLC toafford 12.8 mg (58%) of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)methanesulfonamide28. LCMS-ESI (m/z) calculated for C₂₃H₂₄N₄O₄S: 452.5. found 453.1[M+H]⁺, t_(R)=3.68 min. ¹H NMR (400 MHz, CDCl₃) δ 8.40 (t, J=3.5 Hz,1H), 8.32 (dd, J=8.9, 2.2 Hz, 1H), 7.96 (dd, J=7.7, 1.1 Hz, 1H), 7.67(d, J=7.7 Hz, 1H), 7.38 (t, J=7.7 Hz, 1H), 7.16-7.07 (m, 1H), 4.88-4.70(m, 1H), 4.54 (d, J=8.4 Hz, 1H), 3.19 (dt, J=18.0, 5.9 Hz, 1H), 3.10 (d,J=5.0 Hz, 3H), 3.09-2.95 (m, 1H), 2.14 (qt, J=14.5, 7.3 Hz, 1H),2.06-1.83 (m, 3H), 1.47 (t, J=5.5 Hz, 6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)methanesulfonamide29 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 4.

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-methoxyethanesulfonamide(Compound 31)

Prepared via General Procedure 6 using cyclopropanesulfonyl chloride.LCMS-ESI (m/z) calculated for C₂₅H₂₆N₄O₄S: 478.6. found 479.1 [M+H]⁺,t_(R)=3.84 min.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-methoxyethanesulfonamide(Compound 32)

Prepared via General Procedure 6 using 2-methoxyethanesulfonyl chloride.LCMS-ESI (m/z) calculated for C₂₅H₂₈N₄O₅S: 496.58. found 519.1 [M+Na]⁺,t_(R)=3.83 min. ¹H NMR (400 MHz, CDCl₃) δ 8.41 (d, J=2.2 Hz, 1H), 8.33(dd, J=8.9, 2.2 Hz, 1H), 7.99-7.92 (m, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.37(t, J=7.7 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H), 4.85-4.71 (m, 2H), 4.68-4.60(m, 1H), 3.93-3.76 (m, 2H), 3.47-3.31 (m, 5H), 3.17 (dt, J=18.0, 6.0 Hz,1H), 3.02 (dt, J=18.1, 6.7 Hz, 1H), 2.20-1.82 (m, 4H), 1.48 (d, J=6.1Hz, 6H). Chiral HPLC:(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-methoxyethanesulfonamide32 was eluted with 10% water/MeOH, (Chiral Method 2): 99.98% ee,t_(R)=21.07 min.

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-methoxyethanesulfonamide 33 was prepared in ananalogous fashion from 4. Chiral HPLC: 99.04% ee, t_(R)=18.57 min(Chiral Method 2).

(R)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetate(INT-14)

Prepared using General Procedure 6: To a stirring solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 3 (0.15 g, 0.37 mmol) in DCM (5 mL) were added TEA (76 μL,0.55 mmol) and methyl-2-(chlorosulfonyl)acetate (76 mg, 0.44 mmol).Additional TEA and methyl-2-(chlorosulfonyl)acetate were added to drivethe reaction to completion over 24 h. The crude reaction mixture waspartitioned between DCM and saturated NaHCO₃. The organic layer wasdried over Na₂SO₄, concentrated, and purified by column chromatography(EA/hexanes) to give 0.11 g (57%) of (R)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetateINT-14. LCMS-ESI (m/z) calculated for C₂₅H₂₆N₄O₆S: 510.6. found 511.1[M+H]⁺, t_(R)=3.73 min. ¹H NMR (400 MHz, CDCl₃) δ 8.42 (d, J=2.2 Hz,1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 8.01-7.93 (m, 1H), 7.74 (d, J=7.8 Hz,1H), 7.39 (t, J=7.8 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H), 5.00 (d, J=8.4 Hz,1H), 4.81 (dq, J=18.3, 5.9 Hz, 2H), 4.25-4.00 (m, 2H), 3.83 (s, 3H),3.20 (dt, J=18.1, 5.9 Hz, 1H), 3.12-2.97 (m, 1H), 2.22-2.01 (m, 2H),2.02-1.83 (m, 2H), 1.48 (d, J=6.1 Hz, 6H).

(S)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetateINT-15 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 4.

General Procedure 7. Preparation of Tetrahydronapthalene SulfonamideAcids

To a stirring solution of (R)- or (S)-tetrahydronapthalene sulfonamideester (1 eq) in MeOH (0.2 M) was added 6N NaOH (2 eq) at roomtemperature. The reaction was stirred at room temperature for 6 h. Thecrude reaction was diluted with water, acidified with 1N HCl andextracted with DCM and EA. The organic layer was dried over Na₂SO₄,concentrated, and isolated after preparative HPLC purification.

Compounds 34 and 35 were prepared using General Procedure 7.

(R)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)aceticacid (Compound 34)

Prepared using General Procedure 7: To a stirring solution of (R)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetate INT-14 (0.082 g, 0.16 mmol) in MeOH (1.5 mL) was added 6N NaOH(0.08 mL). The reaction was stirred at room temperature for 6 h. Thecrude reaction was diluted with water, acidified with 1N HCl andextracted with DCM and EA. The organic layer was dried over Na₂SO₄,concentrated, and isolated after preparative HPLC purification to give0.057 g (72%) of(R)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)aceticacid 34. An analytically pure sample was prepared by preparative HPLCpurification. LCMS-ESI (m/z) calculated for C₂₄H₂₆N₄O₆S: 496.5. found520.1 [M+Na]⁺, t_(R)=3.47 min.

(S)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetic acid 35 was prepared in an analogous fashion from(S)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetateINT-15.

General Procedure 8. Preparation of Tetrahydronapthalene SulfonamideAlcohols

To a stirring solution of either (R)- or (S)-tetrahydronapthalenesulfonamide ester (1 eq) in THF (0.06 M) was added sodium borohydride(2.5 eq) at room temperature. The reaction mixture was heated to 75° C.and methanol (1 eq) was added dropwise. After 1 h, the reaction wascooled and concentrated and purified by preparative HPLC.

Compounds 36 and 37 were prepared using General Procedure 8.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-hydroxyethanesulfonamide(Compound 37)

Prepared using General Procedure 8: To a stirring solution of (R)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetateINT-14 (0.025 g, 0.05 mmol) in THF (25 mL) was added sodium borohydride(0.05 g, 0.12 mmol) at room temperature. The reaction was heated to 75°C. and methanol (0.02 mL, 0.05 mmol) was added. After 1 h, the reactionwas cooled and concentrated and purified by preparative HPLC to give16.0 mg (66%) of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-hydroxyethanesulfonamide37. LCMS-ESI (m/z) calculated for C₂₄H₂₆N₄O₅S: 482.6. found 505.1[M+Na]⁺, t_(R)=3.46 min. ¹H NMR (400 MHz, CDCl₃) δ 8.39 (dd, J=5.4, 2.4Hz, 1H), 8.35-8.25 (m, 1H), 7.95 (dt, J=7.7, 3.9 Hz, 1H), 7.67 (d, J=7.5Hz, 1H), 7.43-7.32 (m, 1H), 7.12 (t, J=7.5 Hz, 1H), 4.80 (m, 3H), 4.12(t, J=5.2 Hz, 2H), 3.46-3.28 (m, 2H), 3.17 (dt, J=18.0, 5.9 Hz, 1H),3.02 (dt, J=18.1, 6.8 Hz, 1H), 2.68 (s, 1H), 2.12 (m, 1H), 2.07-1.82 (m,3H), 1.47 (d, J=6.1 Hz, 6H). ¹³C NMR (101 MHz, CDCl₃) δ 172.82, 169.22,162.81, 137.73, 136.90, 134.12, 133.89, 132.15, 130.28, 126.46, 126.33,116.65, 115.22, 113.57, 103.93, 72.78, 57.32, 56.17, 52.54, 30.60,28.06, 21.72, 19.04. Chiral HPLC:(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-hydroxyethanesulfonamide 37 was eluted with 15% water/MeOH, (Chiral Method 2): 99.82%ee, t_(R)=22.23 min.

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-hydroxyethanesulfonamide36 was prepared in an analogous fashion from (S)-methyl2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetateINT-15. Chiral HPLC: 91.7% ee, t_(R)=19.83 min (Chiral Method 2).

General Procedure 9. Preparation of Tetrahydronapthalene SulfonamideAmides

To a stirring solution of either (R)- or (S)-tetrahydronapthalenesulfonamide acid (1 eq) in DMF (0.25 M) were added EDC andN-hydroxybenzotriazole. After 5 min, the amine was added and thereaction mixture was stirred 18 h at room temperature. The crudereaction was diluted with NaHCO₃ added extracted with EA. The combinedorganic layers were dried over Na₂SO₄, and purified by preparative HPLC.

Compounds 38-43 were prepared using General Procedure 9.

(R)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)-N,N-dimethylacetamide(Compound 40)

Prepared using General Procedure 9: To a stirring solution of(R)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)acetic acid 34 (15 mg, 0.05 mmol) in DMF (0.5 mL) was addedN-hydroxybenzotriazole (6.1 mg, 0.05 mmol) and EDC (8.7 mg, 0.05 mmol).After 5 min, dimethylamine (40 wt % solution in THF, 50 μL, 0.09 mmol)was added and the reaction mixture was stirred 18 h at room temperature.The crude reaction was diluted with sat NaHCO₃ and extracted with EA.The combined organic layers were dried over Na₂SO₄ and purified bypreparative HPLC to give 4.41 mg (28%) of(R)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)-N,N-dimethylacetamide40. LCMS-ESI (m/z) calculated for C₂₆H₂₉N₅O₅S: 523.6. found 546.2[M+Na]⁺, t_(R)=3.58 min. ¹H NMR (400 MHz, CDCl₃) δ 8.41 (d, J=2.1 Hz,1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 7.96 (dd, J=7.7, 1.2 Hz, 1H), 7.82(d, J=7.5 Hz, 1H), 7.38 (t, J=7.8 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H), 5.36(t, J=17.3 Hz, 1H), 4.88-4.73 (m, 2H), 4.27 (d, J=14.6 Hz, 1H), 4.07 (d,J=14.6 Hz, 1H), 3.24-3.09 (m, 4H), 3.09-2.97 (m, 4H), 2.23-2.08 (m, 2H),2.10-1.84 (m, 2H), 1.47 (d, J=6.1 Hz, 6H). ¹³C NMR (101 MHz, CDCl₃) δ172.75, 169.37, 163.15, 162.77, 137.79, 136.92, 134.13, 133.90, 132.50,130.18, 126.36, 126.17, 116.79, 115.25, 113.55, 103.96, 72.74, 55.46,53.05, 38.22, 35.98, 29.84, 28.10, 21.73, 19.10.

((S)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)-N,N-dimethylacetamide41 was prepared in an analogous fashion from(S)-2-(N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl) acetic acid 35.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)ethenesulfonamideINT-16

To a stirred solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 3 (100 mg, 0.24 mmol) in DCM (5 mL) at 0° C. were addedTEA (170 μL, 1.2 mmol) and 2-chloroethanesulfonyl chloride (76 μL, 0.73mmol). The reaction mixture was warmed to room temperature and stirredfor 30 min. The solvent was removed and the product was purified bychromatography (EA/hexane) to give 83.0 mg (75%) of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)ethenesulfonamideINT-16 as a white solid. LCMS-ESI (m/z) calculated for C₂₄H₂₄N₄O₄S:464.5. found 465.1 [M+H]⁺, t_(R)=3.83 min. ¹H NMR (400 MHz, CDCl₃) δ8.41 (d, J=2.2 Hz, 1H), 8.32 (dd, J=8.9, 2.2 Hz, 1H), 7.96 (dd, J=7.7,1.2 Hz, 1H), 7.62 (t, J=11.0 Hz, 1H), 7.38 (t, J=7.7 Hz, 1H), 7.12 (d,J=9.0 Hz, 1H), 6.66 (dt, J=26.0, 13.0 Hz, 1H), 6.38 (d, J=16.5 Hz, 1H),6.03 (dd, J=10.1, 5.2 Hz, 1H), 4.86-4.73 (m, 1H), 4.68-4.57 (m, 1H),4.50 (d, J=8.3 Hz, 1H), 3.18 (dt, J=18.1, 5.8 Hz, 1H), 3.02 (dt, J=18.1,6.8 Hz, 1H), 2.15-1.96 (m, 2H), 1.97-1.79 (m, 2H), 1.48 (d, J=6.1 Hz,6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)ethenesulfonamideINT-17 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 4.

General Procedure 10. Preparation of Tetrahydronaphthalene Sulfonamidesvia Michael Addition

To a stirring solution of either the (R)- or (S)-tetrahydronapthalenevinyl sulfonamide (1 eq) in DMF (0.1M) were added TEA (5 eq) and theappropriate amine (5 eq). The reaction mixture was stirred at roomtemperature for 18 h. The products were purified by preparative HPLC.

Compounds 44-47 were prepared using General Procedure 10.

(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-(dimethylamino)ethanesulfonamide(Compound 44)

Prepared using General Procedure 10. To a solution of((R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)ethenesulfonamide INT-16 (40 mg, 0.09 mmol) in DMF (1.0 mL) was added 2Nmethylamine in THF (0.22 mL, 0.43 mmol) and the reaction mixture wasstirred at room temperature for 18 h. The crude product was purified bypreparative HPLC to give 24.6 mg (54%) of the TFA salt of(R)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-(dimethylamino)ethanesulfonamide44 as a white solid. LCMS-ESI (m/z) calculated for C₂₆H₃₁N₅O₄S: 509.6.found 510.2 [M+H]⁺, t_(R)=2.61 min. ¹H NMR (400 MHz, CDCl₃) 8.41-8.32(m, 1H), 8.33-8.26 (m, 1H), 7.92 (t, J=6.9 Hz, 1H), 7.61 (t, J=7.2 Hz,1H), 7.35 (dd, J=14.5, 7.1 Hz, 1H), 7.11 (d, J=9.1 Hz, 1H), 4.86-4.64(m, 2H), 3.61 (ddt, J=27.2, 13.7, 7.8 Hz, 4H), 3.23-3.06 (m, 1H),3.10-2.91 (m, 1H), 2.93 (d, J=30.3 Hz, 6H), 2.09 (ddd, J=28.4, 16.5,12.3 Hz, 1H), 1.95 (ddd, J=15.1, 8.3, 3.5 Hz, 3H), 1.46 (t, J=6.0 Hz,6H).

(S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)-2-(dimethylamino)ethanesulfonamide 45 was prepared in analogous fashion from((S)—N-(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)ethenesulfonamideINT-17.

(R)-2-isopropoxy-5-(3-(5-((2-(methylsulfonyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile(Compound 48)

To a solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 3 (20 mg, 0.05 mmol) in DMA (0.5 mL) was added TEA (136μL, 0.97 mmol) and methylvinylsulfone (52 mg, 0.5 mmol). The reactionwas heated to 80° C. for 24 h. The crude reaction mixture was purifiedby preparative HPLC to give(R)-2-isopropoxy-5-(3-(5-((2-(methylsulfonyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile48. LCMS-ESI (m/z) calculated for C₂₅H₂₈N₄O₄S: 480.6. found 481.2[M+H]⁺, t_(R)=2.58 min. ¹H NMR (400 MHz, CDCl₃) δ 8.37 (t, J=9.1 Hz,1H), 8.31 (dd, J=8.9, 2.2 Hz, 1H), 8.08 (d, J=7.1 Hz, 1H), 7.60 (d,J=7.6 Hz, 1H), 7.42 (t, J=7.7 Hz, 1H), 7.12 (d, J=9.1 Hz, 1H), 4.85-4.73(m, 1H), 4.53 (t, J=5.0 Hz, 1H), 3.63 (dd, J=15.3, 4.0 Hz, 2H),3.61-3.50 (m, 2H), 3.33-3.17 (m, 1H), 3.18-3.04 (m, 1H), 3.04 (d, J=8.6Hz, 3H), 2.16 (ddd, J=29.6, 18.8, 12.2 Hz, 2H), 2.00 (dd, J=36.4, 18.4Hz, 2H), 1.47 (d, J=6.1 Hz, 6H).

(S)-2-isopropoxy-5-(3-(5-((42-(methylsulfonyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile,compound 49 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrilehydrochloride 4.

(R)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetate(INT-18)

To a stirring solution of(R)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile3 (119 mg, 0.32 mmol) in CH₃CN (5.0 mL) was added methyl bromoacetate(53.5 μL, 0.35 mmol) and K₂CO₃ (138 mg, 1.27 mmol). After stirring for18 h, the mixture was diluted with brine and washed with NaHCO₃. Theorganic layer was dried with Na₂SO₄ and concentrated. The resultingcrude solid was purified by chromatography (MeOH/DCM) to provide 113.1mg (79%) of (R)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetate INT-18. LCMS-ESI (m/z) calculated for C₂₅H₂₆N₄O₄: 446.5. found447.2 [M+H]⁺, t_(R)=2.52 min.

(S)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-19 was prepared in an analogous fashion from(S)-5-(3-(5-amino-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile4.

(S)-methyl2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetate(INT-20)

To a stirred solution of (S)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-19 (128.0 mg, 0.29 mmol) in DCM (6.0 mL) was added Boc anhydride(125.1 mg, 0.57 mmol) and TEA (120 μL, 0.86 mmol). After stirring for 18h, the mixture was concentrated. The resulting crude solid was purifiedby chromatography (EA/hexanes) to provide 119 mg (76%) of (S)-methyl2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-20. LCMS-ESI (m/z) calculated for C₃₀H₃₄N₄O₆: 546.61; no M/Zobserved, t_(R)=4.32 min.

(R)-methyl 2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-21 was prepared in an analogous fashion from (R)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-18.

(S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-hydroxyethyl)carbamate(INT-22)

To a stirring solution of (S)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-20 (35 mg, 0.06 mmol) in THF (6.0 mL) at 75° C. was added sodiumborohydride (6 mg, 0.16 mmol). After stirring for 0.5 h, MeOH (7.7 μL,0.19 mmol) was added and the mixture was heated for an additional 1.5 h.The mixture was concentrated and the resulting solid was purified bychromatography (EA/hexanes) to provide 16 mg (48%) of (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-hydroxyethyl)carbamateINT-22. LCMS-ESI (m/z) calculated for C₂₉H₃₄N₄O₅: 518.6. found 419.2[M-Boc+H]⁺, t_(R)=4.10 min.

(R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-hydroxyethyl)carbamateINT-23 was prepared in prepared in an analogous fashion from (R)-methyl2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-21.

General Procedure 11: Boc Deprotection of Tetrahydronaphthalene Amines

To a stirring solution of Boc protected (R)- or(S)-tetrahydronaphthalene amine in dioxane was added 4N HCl/dioxanes(4-10 eq). The reaction mixture was heated at 50° C. for 18 h. Thereaction mixture was concentrated and the resulting solid was purifiedby preparative HPLC.

Compounds 50-53 were prepared using General Procedure 11.

(R)-5-(3-(5-((2-hydroxyethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile(Compound 50)

Prepared using General Procedure 11. To a stirred solution of(R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-hydroxyethyl)carbamate INT-23 (15 mg, 0.03 mmol) in dioxane (1 mL)was added 4N HCl/dioxanes (116 μL, 0.116 mmol). After heating at 50° C.for 18 h, the mixture was concentrated and the resulting solid waspurified by preparative HPLC to provide 9.53 mg (79%) of(R)-5-(3-(5-((2-hydroxyethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile 50. LCMS-ESI (m/z) calculated for C₂₄H₂₆N₄O₃: 418.5. found419.2 [M+H]⁺, t_(R)=2.52 min. ¹H NMR (400 MHz, CDCl₃) δ 8.39 (d, J=2.2Hz, 1H), 8.32 (dd, J=8.9, 2.2 Hz, 1H), 8.04 (d, J=7.7 Hz, 1H), 7.60 (d,J=7.6 Hz, 1H), 7.39 (t, J=7.7 Hz, 1H), 7.12 (d, J=9.1 Hz, 1H), 4.88-4.70(m, 1H), 4.54 (s, 1H), 3.78 (d, J=12.1 Hz, 2H), 3.62 (s, 2H), 3.24 (dt,J=18.0, 5.6 Hz, 1H), 3.18-2.89 (m, 3H), 2.16 (d, J=5.2 Hz, 2H),2.10-1.75 (m, 2H), 1.47 (d, J=6.1 Hz, 6H).

(S)-5-(3-(5-((2-hydroxyethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile51 was prepared in an analogous fashion from (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-hydroxyethyl)carbamateINT-22.

(S)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid (INT-24)

To a stirring solution of (S)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-20 (93.2 mg, 0.17 mmol) in MeOH (2 ml) was added 10 drops of 1 NNaOH. The mixture was stirred at 50° C. for 2 h, then diluted with H₂Oand neutralized with 1 N HCl. The aqueous solution was extracted withDCM and EA. The combined organic layers were dried over Na₂SO₄ andconcentrated to provide 61 mg (67%) of(S)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-24 as a white solid. LCMS-ESI (m/z) calculated for C₂₉H₃₂N₄O₆:532.6. found 358.1 [M-2-((tert-butoxycarbonyl)amino)acetic acid]⁺,t_(R)=3.97 min.

(R)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-25 was prepared in an analogous fashion from (S)-methyl2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)acetateINT-21.

(S)-2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid (Compound 53)

Prepared using General Procedure 11. A solution of(S)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-24 (30 mg, 0.06 mmol) in 4N HCl/dioxanes (200 μl, 50 mmol) wasstirred at room temperature for 18 h. The mixture was concentrated andthe residue was purified by preparative HPLC to provide 21 mg (67%) of(S)-2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid 53 as the TFA salt. LCMS-ESI (m/z) calculated for C₂₄H₂₄N₄O₄:432.5. found 358.1 [M-2-aminoacetic acid]⁺, t_(R)=2.65 min.

(R)-2-((5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino) acetic acid 52 was prepared in an analogousfashion from(R)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-25.

General Procedure 12: Preparation of Tetrahydronaphthalene Amino Amides

To the Boc-protected (R)- or (S)-tetrahydronaphthalene aminoacid DMFwere added N-hydroxybenzotriazole (2 eq) and EDC (2 eq). After 10 min,the appropriate amine (10 eq) was added and the reaction mixture wasstirred for 18 h at room temperature. The crude reaction mixture wasdiluted with NaHCO₃ and extracted with EA. The combined organic layerswere dried over Na₂SO₄ and purified by preparative HPLC.

Compounds 54 and 55 were prepared using General Procedure 12.

(S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate(INT-26)

Prepared using General Procedure 12. To a stirring solution of(S)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-24 (30 mg, 0.06 mmol) in DMF (0.5 mL) were addedN-hydroxybenzotriazole (15.21 mg, 0.11 mmol) and EDC (21.63 mg, 0.11mmol). After 10 min, pyrrolidine (46 μL, 0.56 mmol) was added and thereaction mixture was stirred 18 h at room temperature. The crudereaction was diluted with sat NaHCO₃ added extracted with EA. Thecombined organic layers were dried over Na₂SO₄ and purified bypreparative HPLC to give 26.9 mg (82%) of (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate.LCMS-ESI (m/z) calculated for C₃₃H₃₉N₅O₅: 585.7. found 358.1[M-tert-butyl (2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate]⁺, t_(R)=4.18min.

(R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamateINT-27 was prepared in an analogous fashion from(R)-2-((tert-butoxycarbonyl)(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)amino)aceticacid INT-25.

(R)-2-isopropoxy-5-(3-(5-((2-oxo-2-(pyrrolidin-1-yl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile(Compound 54)

Prepared using General Procedure 11. A solution of (R)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate INT-27 (18 mg, 0.03 mmol) in4N HCl/dioxanes (1 mL) was stirred at room temperature for 18 h. Thereaction mixture was concentrated and purified by preparative HPLC toprovide 12 mg (68%) of(R)-2-isopropoxy-5-(3-(5-((2-oxo-2-(pyrrolidin-1-yl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile54 as the TFA salt. LCMS-ESI (m/z) calculated for C₂₈H₃₁N₅O₃: 485.6.found 486.2 [M+H]⁺, t_(R)=2.60 min. ¹H NMR (400 MHz, CDCl₃) δ 8.40 (d,J=2.2 Hz, 1H), 8.33 (dd, J=8.9, 2.2 Hz, 1H), 8.07 (d, J=7.6 Hz, 1H),7.77 (d, J=7.6 Hz, 1H), 7.44 (t, J=7.7 Hz, 1H), 7.13 (d, J=9.1 Hz, 1H),4.87-4.68 (m, 2H), 3.98-3.80 (m, 2H), 3.53-3.03 (m, 7H), 2.17 (ddd,J=17.7, 10.4, 5.5 Hz, 3H), 2.04-1.79 (m, 5H), 1.48 (d, J=6.1 Hz, 6H).

(S)-2-isopropoxy-5-(3-(5-((2-oxo-2-(pyrrolidin-1-yl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-1-yl)-1,2,4-oxadiazol-5-yl)benzonitrile55 was prepared in an analogous fashion from (S)-tert-butyl(5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1,2,3,4-tetrahydronaphthalen-1-yl)(2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamateINT-26.

4-bromo-2,3-dihydro-1H-inden-1-ol (INT-28)

To a stirring solution of 4-bromoindanone (3 g, 14.2 mmol) in anhydrousEtOH (30 mL) were added sodium borohydride (0.36 g, 9.5 mmol) and silicagel (2 g) at 0° C. The reaction was stirred at 0° C. for 20 min and wasallowed to stir at room temperature for 2 h. The reaction mixture wasquenched with saturated NaHCO₃ and concentrated to remove EtOH. Theaqueous layer was extracted with EA and the organic phase was dried overMgSO₄. After concentration, the crude product was purified bychromatography (EA/hexane) to yield 4-bromo-2,3-dihydro-1H-inden-1-olINT-28 (2.56 g, 85%) as white solid. LCMS-ESI (m/z) calculated forC₉H₉BrO: 213.1. found 195.0 [M−H₂O]⁺, t_(R)=3.07 min. ¹H NMR (400 MHz,CDCl₃) δ 7.35 (d, J=7.9, 1H), 7.27 (d, J=7.4, 1H), 7.05 (t, J=7.7, 1H),5.23 (t, J=6.2, 1H), 3.00 (ddd, J=16.6, 8.8, 4.6, 1H), 2.84-2.66 (m,1H), 2.45 (dddd, J=13.2, 8.4, 7.0, 4.6, 1H), 1.96-1.70 (m, 2H).

(4-bromo-2,3-dihydro-1H-inden-1-yloxy)(tert-butyl)dimethylsilane(INT-29)

To a solution of 4-bromo-2,3-dihydro-1H-inden-1-ol INT-28 (2.56 g, 12.0mm) in DMF (5 mL) were added TBDMSCl (2.17 g, 14.4 mmol) and imidazole(2 g, 30.0 mmol) and the reaction mixture stirred at room temperatureovernight. The reaction mixture was diluted with water and extractedwith EA. The organic layers were washed with water and brine, and driedover MgSO₄. The crude product was purified by chromatography (EA/hexane)to afford(4-bromo-2,3-dihydro-1H-inden-1-yloxy)(tert-butyl)dimethylsilane INT-29(3.3 g, 84%) as a clear oil. LCMS-ESI (m/z) calculated for C₁₅H₂₃BrOSi:327.3. found 195.0 [M-OTBS]⁺, t_(R)=3.07 min. ¹H NMR (400 MHz, CDCl₃) δ7.20 (d, J=7.8 Hz, 1H), 7.06 (d, J=7.4 Hz, 1H), 6.92 (t, J=7.7 Hz, 1H),5.13 (t, J=7.0 Hz, 1H), 2.85 (ddd, J=16.4, 9.1, 2.9 Hz, 1H), 2.57 (dt,J=16.5, 8.3 Hz, 1H), 2.36-2.17 (m, 1H), 1.76 (dtd, J=12.8, 8.8, 7.1 Hz,1H), 0.83-0.72 (m, 9H), 0.05-−0.06 (m, 6H).

tert-butyldimethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-yloxy)silane(INT-30)

A solution of(4-bromo-2,3-dihydro-1H-inden-1-yloxy)(tert-butyl)dimethylsilane INT-29(50 mg, 0.15 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (42 mg, 0.16mmol), and potassium acetate (45 mg, 0.45 mmol) in anhydrous 1,4-dioxane(2 mL) was degassed by passing N₂ through the solution for 5 min.PdCl₂(dppf).CH₂Cl₂ was then added and the reaction mixture was heated at85° C. overnight. The solvent was removed under vacuum, the residue wasdiluted with EA (10 mL), and filtered through celite to remove solids.The filtrate was washed with water and brine and dried over MgSO₄. Thecrude product was purified by chromatography (EA/hexanes) to affordtert-butyldimethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-yl-oxy)silaneINT-30 (26 mg, 45%) as a white semi-solid. LCMS-ESI (m/z) calculated forC₂₁H₃₅BO₃Si: 374.4. found 245.0 [M-OTBS]⁺, t_(R)=3.07 min. ¹H NMR (400MHz, CDCl₃) δ 7.57-7.43 (m, 1H), 7.21 (dd, J=11.0, 4.2 Hz, 1H),7.08-7.01 (m, 1H), 5.06 (t, J=7.0 Hz, 1H), 3.11 (ddd, J=16.8, 8.9, 3.0Hz, 1H), 2.72 (dt, J=16.8, 8.3 Hz, 1H), 2.22 (dddd, J=12.6, 7.9, 7.1,3.1 Hz, 1H), 1.71 (dtd, J=12.6, 8.8, 7.0 Hz, 1H), 1.21-1.10 (m, 12H),0.81-0.71 (m, 9H), 0.03-−0.07 (m, 6H).

5-(4,5-dihydrooxazol-2-yl)-2-isopropoxybenzonitrile (INT-31)

To a stirring suspension of 3-cyano-4-isopropoxybenzoic acid (1.0 g, 4.8mmol) in DCM (20 mL) was added oxalyl chloride (3.7 g, 29.2 mmol)followed by two drops DMF. The reaction mixture was stirred at 50° C.for 2 h. The mixture was concentrated and the residue re-dissolved inDCM (10 mL). Ethanolamine (0.6 g, 9.7 mmol) and TEA (1.45 g, 14.4 mmol)were added and the reaction mixture was stirred overnight at roomtemperature. The resulting solid was filtered, washed with water, anddried to afford 1.0 g (83%) of3-cyano-N-(2-hydroxyethyl)-4-isopropoxybenzamide which was used in thenext step without purification. LCMS-ESI (m/z) calculated forC₁₃H₁₆N₂O₃: 248.3. found 249.0 [M+H]⁺, t_(R)=2.41 min. ¹H NMR (400 MHz,CDCl₃) δ 8.02-7.79 (m, 2H), 6.97-6.87 (m, 1H), 6.71 (s, 1H), 4.65 (dt,J=12.1, 6.1 Hz, 1H), 3.82-3.70 (m, 2H), 3.56 (dd, J=10.2, 5.5 Hz, 2H),1.96 (d, J=10.0 Hz, 1H), 1.40-1.29 (m, 6H).

3-cyano-N-(2-hydroxyethyl)-4-isopropoxybenzamide was dissolved in DCM(30 mL) and thionyl chloride (1.43 g, 12 mmol) was added at 0° C. Thereaction mixture was stirred at room temperature for 1 h and thenquenched at 0° C. with water (200 μL) and 6N NaOH solution (1 mL). Themixture was stirred for 30 min. The aqueous layers were extracted withDCM and the combined organic extracts were washed with brine and driedover MgSO₄ to afford 570 mg (61% for two steps) of5-(4,5-dihydrooxazol-2-yl)-2-isopropoxybenzonitrile INT-31. LCMS-ESI(m/z) calculated for C₁₃H₁₄N₂O₂: 230.3. found 231.0 [M+H]⁺, t_(R)=2.50min. ¹H NMR (400 MHz, CDCl₃) δ 8.17-7.86 (m, 2H), 6.91 (d, J=8.9 Hz,1H), 4.65 (dt, J=12.2, 6.1 Hz, 1H), 4.37 (dd, J=14.3, 4.9 Hz, 2H), 3.98(t, J=9.5 Hz, 2H), 1.36 (t, J=5.5 Hz, 6H).

5-(5-bromooxazol-2-yl)-2-isopropoxybenzonitrile (INT-32)

A stirring solution of5-(4,5-dihydrooxazol-2-yl)-2-isopropoxybenzonitrile INT-31 (420 mg, 1.82mmol), N-bromosuccinamide (990 mg, 5.56 mmol) and azoisobutyronitrile(14.9 mg, 0.09 mmol) in carbon tetrachloride (20 mL) was heated at 80°C. under N₂ for 18 h. The reaction mixture was cooled to roomtemperature and the solids were removed by filtration. The filtrate waswashed with sodium thiosulfate (20 mL) and brine (20 mL), and dried overMgSO₄. The product was purified by chromatography (EA/hexanes) to afford300 mg (55%) of 5-(5-bromooxazol-2-yl)-2-isopropoxybenzonitrile INT-32as a yellow solid. LCMS-ESI (m/z) calculated for C₁₃H₁₁BrN₂O₂: 307.1.found 309.0 [M+2]⁺, t_(R)=3.79 min. ¹H NMR (400 MHz, CDCl₃) δ 8.19-7.93(m, 2H), 7.08-6.85 (m, 2H), 4.81-4.47 (m, 1H), 1.38 (dd, J=6.6, 3.0 Hz,6H).

General Procedure 13: Coupling of Heterocyclic Bromide to IndanolBoronate

A 20 mL microwave vial was charged sequentially with heterocyclicbromide (1 eq), (R)- (S)- or racemic indanol dioxaborolane (1 eq),DME/H₂O (3:1, 0.05 M) and potassium carbonate (3 eq). The mixture wasdegassed by bubbling N₂ gas through the stirring solution for 10 min.Pd(PPh₃)₄ (0.07 eq) was added and the mixture degassed for additional 2min. The vial was capped and subjected to microwave irradiation at 100°C. until reaction completed (40-60 min). Additional bromide was added ifneeded. The vial was cooled to room temperature, diluted with EA (10×volume), washed with water and brine, dried over MgSO₄, andconcentrated. The crude product was purified by silica gel columnchromatography (EA/hexanes).

5-(5-(1-(tert-butyldimethylsilyloxy)-2,3-dihydro-M-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile(INT-33)

Prepared using General Procedure 13. A 20 mL microwave vial was chargedwith 5-(5-bromooxazol-2-yl)-2-isopropoxybenzonitrile INT-32 (200 mg,0.65 mmol),tert-butyldimethyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-yloxy)silaneINT-30 (243 mg, 0.65 mmol), potassium carbonate (269 mg, 1.95 mmol) anda 3:1 mixture of dimethylethylene glycol/H₂O (10 mL). The reactionmixture was degassed by bubbling N₂ gas through the stirring solutionfor 10 min. Pd(PPh₃)₄ was added and the solution degassed for additional2 min. The vial was subjected to microwave irradiation at 100° C. for 40min. The vial was cooled to 0° C. and the resulting solid obtained wascollected by filtration, washed with ice water, and dried to afford 290mg (94%) of5-(5-(1-(tert-butyldimethylsilyloxy)-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile INT-33 as a light yellow solid.LCMS-ESI (m/z) calculated for C₂₈H₃₄N₂O₃Si: 474.7. found 475.2 [M+H]⁺,t_(R)=5.90 min (Method 1). ¹H NMR (400 MHz, CDCl₃) δ 8.16-7.96 (m, 2H),7.57-7.42 (m, 1H), 7.24-7.12 (m, 3H), 6.90 (t, J=10.4 Hz, 1H), 5.14 (t,J=7.0 Hz, 1H), 4.57 (dt, J=12.3, 6.1 Hz, 1H), 3.04 (ddd, J=16.1, 9.1,3.1 Hz, 1H), 2.78 (dt, J=16.1, 8.1 Hz, 1H), 2.43-2.24 (m, 1H), 1.84(ddd, J=15.8, 12.8, 8.9 Hz, 1H), 1.27 (t, J=5.8 Hz, 6H), 0.86-0.61 (m,9H), 0.06-−0.14 (m, 6H).

5-(5-(1-hydroxy-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile(Compound 56)

To a solution of5-(5-(1-(tert-butyldimethylsilyloxy)-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrileINT-33 (350 mg, 0.737 mmol) in anhydrous THF (2 mL) was added a 1Msolution of tetrabutylammonium fluoride in THF (3.6 mL, 3.6 mmol) at 0°C. The reaction mixture was allowed to stir at room temperature for 16 hbefore quenching with brine (5 mL). The THF was removed under vacuum,the residue was diluted with water (5 mL), and the aqueous layer wasextracted with EA. The combined extracts were washed with brine, driedover MgSO₄, and purified by chromatography to afford 220 mg (63%) of5-(5-(1-hydroxy-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile56 as a light yellow solid. LCMS-ESI (m/z) calculated for C₂₂H₂₀N₂O₃:360.4. found 343.0 [M-OH]⁺, t_(R)=2.30 min. ¹H NMR (400 MHz, CDCl₃) δ8.30 (d, J=2.2 Hz, 1H), 8.26 (dd, J=8.9, 2.2 Hz, 1H), 7.75 (d, J=7.6 Hz,1H), 7.48 (d, J=7.3 Hz, 1H), 7.42 (t, J=7.6 Hz, 1H), 7.10 (d, J=8.9 Hz,1H), 5.35 (d, J=4.8 Hz, 1H), 4.78 (dt, J=12.2, 6.1 Hz, 1H), 3.30 (ddd,J=16.4, 8.7, 4.8 Hz, 1H), 3.13-2.94 (m, 1H), 2.64 (dddd, J=13.3, 8.4,7.1, 4.8 Hz, 1H), 2.17-2.08 (m, 1H), 1.86 (s, 1H), 1.60 (s, 1H), 1.46(dd, J=13.9, 6.0 Hz, 6H).

General Procedure 14. Preparation of Indane Amines via ChlorideDisplacement

To a stirring solution of indane alcohol (1 eq) in DCM (1 mL) was addedthionyl chloride (2 eq.) at 0° C. The reaction mixture was stirred atroom temperature for 3 h. The solvent was evaporated and the crudechloride re-dissolved in dimethyl acetamide (1 mL). Diisopropylethylamine (3 eq.) and the appropriate amine (3 eq.) were added and thereaction mixtures were stirred at 70° C. overnight. The reactionmixtures were quenched with water (200 μL) and purified by preparativeHPLC.

Compounds 57, 58, and 61-64 were prepared using General Procedure 14.

5-(5-(1-(2-hydroxyethylamino)-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile(Compound 57)

Prepared using General Procedure 14. To a stirring solution of5-(5-(1-hydroxy-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile56 (50 mg, 0.1 mmol) in DCM (3 mL) was added thionyl chloride (25 mg,0.21 mmol) at 0° C. The reaction mixture was stirred at room temperaturefor 3 h. The solvent was evaporated and the crude chloride re-dissolvedin dimethyl acetamide (3 mL). Isopropyl ethylamine (40.8 mg, 0.316 mmol)and ethanolamine (19.3 mg, 0.31 mmol) were added and the reactionmixture heated at 70° C. overnight. The reaction mixture was quenchedwith NaHCO₃ and extracted with EA. The combined organic extracts werewashed with brine and then dried over MgSO₄. The product was purified bychromatography (10% MeOH/DCM) to afford 25 mg (60%) of5-(5-(1-(2-hydroxyethylamino)-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile57. LCMS-ESI (m/z) calculated for C₂₄H₂₅N₃O₃: 403.5. found 404.1 [M+H]⁺,t_(R)=2.41 min. ¹H NMR (400 MHz, DMSO) δ 8.18 (t, J=2.3 Hz, 1H), 8.08(dd, J=9.0, 2.3 Hz, 1H), 7.70 (d, J=7.7 Hz, 1H), 7.44 (d, J=17.4 Hz,1H), 7.42-7.32 (m, 1H), 7.30-7.11 (m, 2H), 4.70 (dt, J=12.2, 6.1 Hz,2H), 4.39 (s, 1H), 3.40 (t, J=5.0 Hz, 2H), 3.18-2.95 (m, 2H), 2.93-2.75(m, 1H), 2.73-2.54 (m, 2H), 2.38-2.16 (m, 1H), 1.98-1.78 (m, 1H), 1.15(d, J=6.0 Hz, 6H). ¹³C NMR (101 MHz, CDCl₃) δ 161.35, 159.17, 151.04,146.60, 139.78, 132.16, 127.43, 125.59, 125.07, 123.99, 120.49, 116.10,113.90, 103.77, 72.60, 62.95, 61.51, 48.70, 33.27, 31.29, 29.91, 22.02.

5-(5-(1-((R)-1-hydroxypropan-2-ylamino)-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrile(Compound 58)

Prepared using General Procedure 14. LCMS-ESI (m/z) calculated for:C₂₅H₂₇N₃O₃: 417.5. found 418.4 [M+H]⁺, t_(R)=2.49 min.

(R)—N—((R)-4-bromo-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamide(INT-34)

To a stirring solution of 4-bromo-2,3-dihydro-1H-inden-1-one (5.0 g,23.6 mmol) and (R)-2-methylpropane-2-sulfinamide (3.15 g, 26.0 mmol) intoluene (40 mL) was added titanium tetraethoxide (8.1 g, 35.5 mmol) andthe reaction mixture was heated at 60° C. for 18 h under N₂. To thismixture was added THF (40 mL) and the resulting solution was cooled to−78° C. Sodium borohydride (3.5 g, 94.7 mmol) was added in one portion.The reaction mixture was stirred at −78° C. for 15 min, and then warmedto room temperature and stirred at this temperature for 2 h. Thereaction mixture was cooled to 0° C. before quenching with brine andsodium potassium tartrate. EA was added and the mixture was stirred atroom temperature overnight during which time Ti salts precipitated. Theorganic layers were decanted, and washed successively with saturatedNH₄Cl, water, and brine. The organic layers were dried over MgSO₄,filtered through a pad of MgSO₄, and concentrated to produce(R)—N—((R)-4-bromo-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-34 as a solid (3.14 g, 42%) which was used in the next step withoutpurification. LCMS-ESI (m/z) calculated C₁₃H₁₈BrNOS: 317.3. found 318.0[M+H]⁺, t_(R)=3.59 min. ¹H NMR (400 MHz, CDCl₃) δ 7.46 (d, J=7.5, 1H),7.34 (d, J=7.9, 1H), 7.05 (t, J=7.7, 1H), 4.96-4.77 (m, 1H), 3.39 (d,J=6.8, 1H), 3.06-2.86 (m, 1H), 2.82-2.60 (m, 1H), 2.50-2.29 (m, 1H),2.05-1.81 (m, 1H), 1.16 (s, 9H).

(S)—N—((S)-4-bromo-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-35 can be made in an analogous fashion using(S)-2-methylpropane-2-sulfinamide.

(R)-4-bromo-2,3-dihydro-1H-inden-1-amine (INT-36)

To crude(R)—N—((R)-4-bromo-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-34 (3.14 g, 9.9 mol) in MeOH (10 mL) was added 4N HCl in dioxane(7.5 mL, 30 mmol) and the resulting yellow suspension was stirred atroom temperature for 2 h. The crude reaction mixture diluted with MeOH(5 mL), cooled to 0° C., and filtered to remove Ti by-products. Thefiltrate was concentrated and the resulting solid refluxed inacetonitrile (60 mL) for 30 min and then cooled to 0° C. The resultingwhite solid was collected to produce the HCl salt of(R)-4-bromo-2,3-dihydro-1H-inden-1-amine INT-36 (1.55 g, 63%) which wasused in the next step without purification. LCMS-ESI (m/z) calculatedfor C₉H₁₀BrN: 212.1. found 197.0 [M-NH]⁺, t_(R)=0.75 min. ¹H NMR (400MHz, DMSO) δ 8.60 (s, 1H), 7.67 (d, J=7.5 Hz, 1H), 7.57 (d, J=7.9 Hz,1H), 7.39-7.07 (m, 1H), 4.81 (dd, J=7.9, 5.6 Hz, 1H), 3.25-2.64 (m, 3H),2.59-2.32 (m, 1H), 2.21-1.69 (m, 1H).

(S)-4-bromo-2,3-dihydro-1H-inden-1-amine INT-37 can be made in ananalogous fashion from(S)—N—((S)-4-bromo-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-35.

(R)-tert-butyl 4-bromo-2,3-dihydro-1H-inden-1-ylcarbamate (INT-38)

To crude (R)-4-bromo-2,3-dihydro-1H-inden-1-amine HCl INT-36 (1.55 g,6.2 mmol) in DCM (10 mL) at 0° C. was added TEA (1.38 g, 13.7 mmol)followed by Boc anhydride (1.49 g, 6.8 mmol) and the reaction mixturestirred at room temperature overnight. The reaction mixture was washedwith brine, and the organic layers were dried over MgSO₄ and filtered.The product was purified by chromatography (EA/hexanes) to afford(R)-tert-butyl 4-bromo-2,3-dihydro-1H-inden-1-ylcarbamate INT-38 (1.63g, 84%) as an off-white solid. LCMS-ESI (m/z) calculated forC₁₄H₁₈BrNO₂: 312.20. found 197.0 [M-NHBoc]⁺, t_(R)=3.97 min. ¹H NMR (400MHz, CDCl₃) δ 7.31 (d, J=7.9 Hz, 1H), 7.23-7.13 (m, 1H), 7.02 (t, J=7.7Hz, 1H), 5.30-5.07 (m, 1H), 4.69 (d, J=7.5 Hz, 1H), 2.93 (ddd, J=16.5,9.0, 3.4 Hz, 1H), 2.75 (dt, J=16.5, 8.2 Hz, 1H), 2.60-2.43 (m, 1H), 1.73(dq, J=13.1, 8.4 Hz, 1H), 1.41 (s, 9H).

(S)-tert-butyl 4-bromo-2,3-dihydro-1H-inden-1-ylcarbamate INT-39 can bemade in an analogous fashion from(S)-4-bromo-2,3-dihydro-1H-inden-1-amine INT-37.

(R)-tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ylcarbamate(INT-40)

A solution of (R)-tert-butyl 4-bromo-2,3-dihydro-1H-inden-1-ylcarbamateINT-38 (300 mg, 0.96 mmol) and4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (268 mg, 1.0mmol), potassium acetate (283 mg, 2.88 mmol) in anhydrous 1,4-dioxane (5mL) was degassed by passing N₂ through the solution for 5 min.PdCl₂(dppf).DCM (157 mg, 0.19 mmol) was added and the reaction mixturewas heated at 85° C. overnight. The solvent was removed under vacuum andthe residue dissolved in EA (10 mL) and filtered through celite toremove the solids. The filtrate was washed with water and brine, driedover MgSO₄, and purified by chromatography (EA/hexanes) to afford(R)-tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-40 (265 mg, 77%) as white semi-solid. LCMS-ESI (m/z) calculated forC₂₀H₃₀BNO₄: 359.3. found 383.0 [M+Na]⁺, t_(R)=4.26 min. ¹H NMR (400 MHz,CDCl₃) δ 7.71 (d, J=7.3 Hz, 1H), 7.42 (t, J=7.9 Hz, 1H), 7.24 (dd,J=9.7, 5.2 Hz, 1H), 5.19 (dd, J=15.9, 7.9 Hz, 1H), 4.72 (d, J=8.5 Hz,1H), 3.28 (ddd, J=17.0, 8.8, 3.6 Hz, 1H), 2.99 (dt, J=16.8, 8.4 Hz, 1H),2.69-2.44 (m, 1H), 1.77 (ddd, J=16.4, 12.8, 8.6 Hz, 1H), 1.51 (s, 9H),1.39-1.31 (m, 12H).

(S)-tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-41 can be made in an analogous fashion from (S)-tert-butyl4-bromo-2,3-dihydro-1H-inden-1-ylcarbamate INT-39.

(R)-tert-butyl4-(2-(3-cyano-4-isopropoxyphenyl)oxazol-5-yl)-2,3-dihydro-1H-inden-1-ylcarbamate(INT-42)

Prepared using General Procedure 13. A 20 mL microwave vial was chargedwith (R)-tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-32 (58.4 mg, 0.16 mmol),5-(5-bromooxazol-2-yl)-2-isopropoxybenzonitrile INT-40 (50 mg, 0.16mmol), potassium carbonate (68 mg, 0.5 mmol) and a 3:1 mixture ofdimethylethylene glycol/H₂O (2 mL). The reaction mixture was degassed bybubbling N₂ gas through the stirring solution for 10 min. Pd(PPh₃)₄((3.9 mg, 0.004 mmol) was added and the solution degassed for additional2 min. The vial was subjected to microwave irradiation at 100° C. for 30min. The solvent was removed and the residue dissolved in EA (10 mL),washed with brine, and then dried over MgSO₄. The product was purifiedby chromatography (EA/hexanes) to afford 50 mg (67%) of (R)-tert-butyl4-(2-(3-cyano-4-isopropoxyphenyl)oxazol-5-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-42 as an off-white solid. LCMS-ESI (m/z) calculated for C₂₇H₂₉N₃O₄:459.5. found 460.2 [M+H]⁺, t_(R)=4.1 min. ¹H NMR (400 MHz, CDCl₃) δ8.32-8.03 (m, 2H), 7.60 (dd, J=8.6, 4.1 Hz, 1H), 7.32-7.22 (m, 3H), 7.00(d, J=8.9 Hz, 1H), 5.19 (dd, J=15.5, 7.5 Hz, 1H), 4.82-4.56 (m, 2H),3.12 (ddd, J=16.3, 9.0, 3.5 Hz, 1H), 2.95 (dt, J=16.3, 8.1 Hz, 1H),2.70-2.51 (m, 1H), 1.83 (dq, J=13.1, 8.2 Hz, 1H), 1.43 (s, 9H),1.41-1.35 (m, 6H).

(S)-tert-butyl4-(2-(3-cyano-4-isopropoxyphenyl)oxazol-5-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-43 can be made in an analogous fashion from (S)-tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-41.

(R)-5-(5-(1-amino-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxy-benzonitrilehydrochloride (Compound 59)

To a stirring solution of (R)-tert-butyl4-(2-(3-cyano-4-isopropoxyphenyl)oxazol-5-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-42 (48 mg, 0.1 mmol) in 1,4-dioxane (1 mL) was added a 4N HClsolution in 1,4-dioxane (1 mL). The reaction mixture was heated at55-65° C. for 48 h. The cooled reaction mixture was diluted with Et₂O(10 mL). The resulting solid was collected and dried under high vacuumto yield 32 mg (78%) of(R)-5-(5-(1-amino-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrilehydrochloride 59 as a white solid. LCMS-ESI (m/z) calculated forC₂₂H₂₁N₃O₂: 359.4. found 343.1 [M-NH₂]⁺, t_(R)=2.40 min. ¹H NMR (400MHz, DMSO) δ 8.55 (br s, 2H), 8.43 (dd, J=6.5, 2.4 Hz, 1H), 8.32 (ddd,J=6.7, 6.1, 2.9 Hz, 1H), 8.00 (t, J=13.5 Hz, 1H), 7.72 (s, 1H), 7.66 (d,J=7.5 Hz, 1H), 7.49 (dd, J=8.5, 6.3 Hz, 2H), 4.93 (dt, J=12.1, 6.0 Hz,1H), 4.81 (s, 1H), 3.43-3.25 (m, 1H), 3.23-3.04 (m, 1H), 2.67-2.55 (m,1H), 2.11 (ddd, J=14.2, 9.0, 5.9 Hz, 1H), 1.36 (dd, J=13.8, 7.0 Hz, 6H).

(S)-5-(5-(1-amino-2,3-dihydro-1H-inden-4-yl)oxazol-2-yl)-2-isopropoxybenzonitrilehydrochloride INT-44 can be made in an analogous fashion from(S)-tert-butyl4-(2-(3-cyano-4-isopropoxyphenyl)oxazol-5-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-43.

1-oxo-2,3-dihydro-1H-indene-4-carbonitrile (INT-45)

To a stirring solution of 4-bromo-2,3-dihydro-1H-inden-1-one (100.0 g,0.48 mol) in 150 mL of 1-methy-2-pyrrolidine (NMP) was added zinccyanide (111.8 g, 0.95 mol) and tetrakis(triphenylphosphine)palladium[Pd(PPh₃)₄] (2.75 g, 0.024 mol). The solution was degassed with N₂ andthe reaction mixture heated at 95° C. for 7 h. Upon cooling, thereaction mixture was poured onto ice water (3.5 L). The compound andinorganic Zn salts precipitated. The solid was collected and partitionedbetween DCM and water. The organic layers were filtered to remove the Znsalts, and the filtrate was concentrated and crystallized from a 4:1mixture of EtOH and MeOH (400 mL) to give 45.5 g (60%) of1-oxo-2,3-dihydro-1H-indene-4-carbonitrile INT-45 as a light yellowsolid. LCMS-ESI (m/z) calculated for C₁₀H₇NO: 157.2. found 158.1 [M+H]⁺,t_(R)=2.67 min. ¹H NMR (400 MHz, CDCl₃) δ 8.00-7.90 (m, 1H), 7.86 (dd,J=7.5, 1.1, 1H), 7.50 (t, J=7.6, 1H), 3.40-3.19 (m, 2H), 2.90-2.61 (m,2H). ¹³C NMR (101 MHz, CDCl₃) δ 204.70, 157.90, 138.38, 137.88, 128.44,128.28, 116.31, 111.70, 36.01, 25.49.

(±)-1-hydroxy-2,3-dihydro-1H-indene-4-carbonitrile (INT-46)

To a stirring suspension of 1-oxo-2,3-dihydro-1H-indene-4-carbonitrileINT-45 (1.2 g, 7.64 mmol) and silica gel (catalytic) in EtOH at 0° C.was added NaBH₄ (237.2 mg, 7.64 mmol). The reaction was allowed to warmto room temperature and stirred for 2 h. The solvent was removed underreduced pressure, and the product was purified by chromatography(EA/hexane) to afford 1.02 g (82%) of1-hydroxy-2,3-dihydro-1H-indene-4-carbonitrile INT-46 as a white solid.LCMS-ESI (m/z) calculated for C₁₀H₉NO; 159.2. found 160.1 [M+H]⁺,t_(R)=2.39 min.

N,1-dihydroxy-2,3-dihydro-1H-indene-4-carboximidamide (INT-47)

Prepared using General Procedure 1. To hydroxylamine hydrochloride (0.87g, 12.5 mmol) and sodium carbonate (1.32 g, 12.5 mmol) in EtOH (20 mL)was added 1-hydroxy-2,3-dihydro-1H-indene-4-carbonitrile INT-46 (1.80 g,11.3 mmol) in one portion and the solution was heated to reflux. After16 h, the reaction was cooled and filtered to remove the solids. TheEtOH was removed and the compound was purified by chromatography(MeOH/DCM) to give 1.74 g (90%) ofN,1-dihydroxy-2,3-dihydro-1H-indene-4-carboximidamide INT-47 as a whitefoam. LCMS-ESI (m/z) calculated for C₁₀H₁₂N₂O₂: 192.1. found: 193.1[M+H]⁺, t_(R)=0.56 min. ¹H NMR (400 MHz, MeOD) δ 10.30 (s, 1H), 9.97 (s,1H), 7.72-7.58 (m, 1H), 7.46-7.37 (m, 2H), 5.22 (t, J=6.5, 1H),3.17-3.03 (m, 1H), 2.99-2.83 (m, 1H), 2.49 (dddd, J=11.4, 8.0, 7.0, 4.4,1H), 2.02-1.88 (m, 1H).

4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol(Compound 60)

Prepared using General Procedure 2. A solution of2-(3,4-diethoxyphenyl)acetic acid (180.0 mg, 0.80 mmol) in DMF (3 mL)was treated with HOBt (197.8 mg, 1.46 mmol) and EDC (207.3 mg, 1.08mmol) at room temperature. The reaction was stirred for 2 h until thecomplete formation of the HOBt-acid complex.N-1-dihydroxy-2,3-dihydro-1H-indene-4-carboximidamide INT-47 (185.1 mg,0.96 mmol) was added and the mixture was stirred at room temperature for2 h and then heated to 80° C. for 16 h. The reaction mixture was dilutedwith NaHCO₃ and extracted with EA. The organic phase was dried overMgSO₄ and crude product was purified by chromatography (EA/hexanes) toproduce4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol60 (190 mg, 62%) as an off-white solid. LCMS-ESI (m/z) calculated forC₂₂H₂₄N₂O₄: 380.1. found 381.1 [M+H]⁺, t_(R)=3.45 min. ¹H NMR (400 MHz,CDCl₃) δ 8.02-7.86 (m, 1H), 7.46 (d, J=7.5 Hz, 1H), 7.29 (t, J=7.6 Hz,1H), 6.81 (ddd, J=21.0, 13.6, 5.1 Hz, 3H), 5.21 (t, J=5.6 Hz, 1H), 4.13(s, 2H), 4.01 (dq, J=14.1, 7.0 Hz, 4H), 3.34 (ddd, J=17.5, 8.7, 4.6 Hz,1H), 3.16-2.92 (m, 1H), 2.53-2.38 (m, 1H), 1.91 (qdd, J=8.7, 6.6, 5.5Hz, 2H), 1.36 (td, J=7.0, 4.6 Hz, 6H).

2-((4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)ethanol(Compound 61)

Prepared using General Procedure 14 using4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol60 and 2-aminoethanol.

(2R)-2-((4(4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)propan-1-ol(Compound 62)

Prepared using General Procedure 14 from4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol60 and (R)-2-aminopropan-1-ol.

4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-N-(2-(methylsulfonyl)ethyl)-2,3-dihydro-1H-inden-1-amine(Compound 63)

Prepared using General Procedure 14 from4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol60 and 2-(methylsulfonyl)ethanamine.

2-((4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)-N-methylethanesulfonamide(Compound 64)

Prepared using General Procedure 14 from4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ol60 and 2-amino-N,N-dimethylethanesulfonamide.

(R)—N-(4-cyano-2,3-dihydro-1H-indene-1-ylidene)-2-methylpropane-2-sulfinamide(INT-48)

To 1-oxo-2,3-dihydro-1H-indene-4-carbonitrile INT-45 (42.5 g, 0.27 mol)and (R)-2-methylpropane-2-sulfinamide (36.0 g, 0.30 mol) in toluene (530mL) was added titanium tetraethoxide (84.1 mL, 92.5 g, 0.40 mol) and thereaction mixture was heated at 60° C. for 12 h under N₂. The crude(R)—N-(4-cyano-2,3-dihydro-1H-indene-1-ylidene)-2-methylpropane-2-sulfinamideINT-48 was used directly in the next experiment. LCMS-ESI (m/z)calculated for C₁₄H₁₆N₂OS: 260.3. found 261.1 [M+H]⁺, t_(R)=3.19 min.

(R)—N—((R)-4-cyano-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamide(INT-49)

To a flask containing the crude suspension of(R)—N-(4-cyano-2,3-dihydro-1H-indene-1-ylidene)-2-methylpropane-2-sulfinamideINT-48 under N₂ was added THF (1.0 L) and the reaction mixture cooled to−78° C. Sodium borohydride (40.9 g, 1.08 mol) was added portion-wiseover 30 mins. (The internal temperature did not rise during theaddition.) The reaction mixture was stirred at −78° C. for 30 mins, halfout of the bath for 30 mins, then warmed to 0° C. over 1 h. The 0° C.reaction mixture was placed in an ice bath and quenched with brine (100mL) followed by saturated sodium potassium tartrate (420 mL) and the Tisalts precipitated. The reaction mixture was diluted with EA (1.5 L) andstirred at room temperature overnight. The organic layers were decantedand washed successively with saturated NH₄Cl, water, and brine. Theorganic layers were dried over MgSO₄ and filtered through a pad ofMgSO₄. The filtrate was concentrated to produce 52.9 g of crude(R)—N—((R)-4-cyano-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-49 as a brown oil, which was used directly in the next step.LCMS-ESI (m/z) calculated for C₁₄H₁₈N₂OS: 262.3. found 263.1 [M+H]⁺,t_(R)=2.99 min. ¹H NMR (400 MHz, CDCl₃) δ 7.89 (d, J=7.7, 1H), 7.56 (t,J=6.8, 1H), 7.36 (t, J=7.7, 1H), 4.97 (q, J=7.5, 1H), 3.50 (d, J=7.6,1H), 3.22 (ddd, J=16.9, 8.8, 3.9, 1H), 3.01 (dt, J=22.4, 6.9, 1H),2.70-2.53 (m, 1H), 2.15-1.95 (m, 1H), 1.33-1.20 (m, 9H).

(R)-1-amino-2,3-dihydro-1H-indene-1-yl)-4-carbonitrile (INT-50)

To crude(R)—N—((R)-4-cyano-2,3-dihydro-1H-inden-1-yl)-2-methylpropane-2-sulfinamideINT-49 (52.9 g, 0.20 mol) in MeOH (200 mL) was added 4N HCl in dioxane(152.0 mL, 0.60 mol) and the resulting yellow suspension was stirred atroom temperature for 1.5 h. The crude reaction mixture was diluted withMeOH (500 mL) and filtered to remove some Ti by-products. The filtratewas concentrated and the resulting solid was refluxed in acetonitrile(500 mL). The resulting white solid was collected to produce 13.0 g (31%over 3 steps) of the HCl salt of(R)-1-amino-2,3-dihydro-1H-indene-1-yl)-4-carbonitrile INT-50. LCMS-ESI(m/z) calculated for C₁₀H₁₀N₂: 158.2. found 142.0 [M-NH₂]⁺, t_(R)=0.84min. ¹H NMR (400 MHz, DMSO) δ 8.61 (s, 3H), 7.96 (d, J=7.7, 1H), 7.83(d, J=7.5, 1H), 7.52 (t, J=7.7, 1H), 4.80 (s, 1H), 3.23 (ddd, J=16.6,8.7, 5.2, 1H), 3.05 (ddd, J=16.6, 8.6, 6.3, 1H), 2.62-2.51 (m, 1H),2.15-2.01 (m, 1H). ¹³C NMR (101 MHz, DMSO) δ 148.09, 141.15, 132.48,130.32, 127.89, 117.27, 108.05, 54.36, 39.08, 29.64. The free base canbe prepared by extraction with 1N NaHCO₃ and DCM. LCMS-ESI (m/z)calculated for C₁₀H₁₀N₂: 158.2. found 142.0 [M-NH₂]⁺, t_(R)=0.83 min. ¹HNMR (400 MHz, CDCl₃) δ 7.52-7.38 (m, 2H), 7.23 (dd, J=17.4, 9.8, 1H),4.35 (t, J=7.6, 1H), 3.11 (ddd, J=16.8, 8.7, 3.2, 1H), 2.89 (dt, J=16.9,8.5, 1H), 2.53 (dddd, J=12.8, 8.1, 7.3, 3.2, 1H), 1.70 (dtd, J=12.8,8.8, 8.0, 1H). ¹³C NMR (101 MHz, DMSO) δ 150.16, 146.67, 130.19, 128.74,127.38, 117.77, 107.42, 56.86, 38.86, 29.14. Chiral HPLC:(R)-1-amino-2,3-dihydro-1H-indene-1-yl)-4-carbonitrile was eluted using5% EtOH in hexanes, plus 0.05% TEA: 95% ee, t_(R)=23.02 min.

The (S)-enantiomer INT-51 was prepared in an analogous sequence (INT-48,INT-49, and INT-50) using (S)-2-methylpropane-2-sulfinamide in the firststep. t_(R) for (S)-enantiomer=20.17 min.

(R)-tert-butyl 4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate (INT-52)

To (R)-1-amino-2,3-dihydro-1H-indene-1-yl)-4-carbonitrile HCl INT-50(11.6 g, 59.6 mmol) in DCM (100 mL) at 0° C. was added TEA (12.0 mL,131.0 mmol). To the resulting solution was added a solution of Bocanhydride (14.3 g, 65.6 mmol) in DCM (30 mL) and the reaction mixturestirred at room temperature for 1.5 h. The reaction mixture was washedwith brine, and the organic layers were dried over MgSO₄ and filtered.Additional DCM was added to a total volume of 250 mL and Norit (4.5 g)was added. The product was refluxed for 15 mins and the hot mixturefiltered through a pad of celite/silica. The filtrate was concentratedand recrystallized from EA (50 mL) and hexane (150 mL) to produce 12.93g (84%) of (R)-tert-butyl 4-cyano-2,3-dihydro-1H-inden-1-ylcarbamateINT-52 as an off-white solid. LCMS-ESI (m/z) calculated for C₁₅H₁₈N₂O₂:258.3. found 281.1 [M+Na]⁺, t_(R)=3.45 min. Elemental Analysisdetermined for C₁₅H₁₈N₂O₂; C calculated=69.74%. found=69.98%. Hcalculated=7.02%. found=7.14%. N calculated=10.84%. found=10.89%. ¹H NMR(400 MHz, CDCl₃) δ 7.64-7.49 (m, 2H), 7.34 (dt, J=7.7, 3.8, 1H),5.36-5.20 (m, 1H), 4.78 (d, J=6.8, 1H), 3.20 (ddd, J=16.9, 8.9, 3.3,1H), 3.02 (dt, J=25.4, 8.4, 1H), 2.82-2.53 (m, 1H), 1.88 (dq, J=13.2,8.6, 1H), 1.55-1.44 (m, 9H). ¹³C NMR (101 MHz, DMSO) δ 155.52, 146.68,146.32, 130.89, 128.70, 127.63, 117.51, 107.76, 77.98, 55.09, 31.88,29.11, 28.19. Chiral HPLC: (R)-tert-butyl4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate was eluted using 2.5% EtOH inhexanes: >99.9% ee, t_(R)=19.36 min.

The (S)-enantiomer INT-53 was prepared in an analogous fashion using(S)-1-amino-2,3-dihydro-1H-indene-1-yl)-4-carbonitrile HCl INT-51. t_(R)for (S)-enantiomer=28.98 min.

(R)-tert-butyl4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-ylcarbamate (INT-54)

Prepared using General Procedure 1. To (R)-tert-butyl4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate INT-52 (15.0 g, 58.2 mmol) inEtOH (100 mL) was added hydroxylamine hydrochloride (12.1 g, 174.2 mmol)and TEA (17.6 mL, 174.2 mmol) and the reaction mixture was heated at 85°C. for 2 h. The solvents were removed and the resulting white solid waspartitioned between water and DCM. The organic layers were dried overNa₂SO₄, concentrated, and recrystallized from isopropanol (50 mL) toafford 14.4 g (85%) of (R)-tert-butyl4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-ylcarbamate INT-54 aswhite crystalline solid. LCMS-ESI (m/z) calculated for C₁₅H₂₁N₃O₃:291.4. found 292.1 [M+H]⁺, t_(R)=2.04 min. ¹H NMR (400 MHz, DMSO) δ 9.53(s, 1H), 7.38-7.32 (m, 1H), 7.32-7.12 (m, 3H), 5.68 (s, 2H), 4.97 (q,J=8.5, 1H), 3.07 (ddd, J=16.6, 8.7, 2.6, 1H), 2.86 (dt, J=16.8, 8.4,1H), 2.30 (ddd, J=12.6, 7.6, 3.6, 1H), 1.75 (dq, J=12.3, 9.0, 1H), 1.44(s, 9H).

(S)-tert-butyl4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-ylcarbamate INT-55 wasprepared in an analogous fashion from (R)-tert-butyl4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate INT-53.

(R)-tert-butyl4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylcarbamate(INT-56)

Prepared using General Procedure 2. A solution of2-(3,4-diethoxyphenyl)acetic acid (150.0 mg, 0.67 mmol) in DMF (3 mL)was treated with HOBt (164.8 mg, 1.22 mmol) and EDC (172.7 mg, 0.9 mmol)at room temperature. The reaction was stirred for 2 h until the completeformation of the HOBt-acid complex. (R)-tert-butyl4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-ylcarbamate INT-54(233.8 mg, 0.8 mmol) was added and stirred at room temperature for 2 hand then mixture was heated to 80° C. for 16 h. The reaction was dilutedwith NaHCO₃ (10 mL) and extracted with EA (3×10 ml). The organic phasewas dried over MgSO₄ and the crude product was purified by achromatography (EA/hexanes) to produce (R)-tert-butyl4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-56 (187 mg, 58%) as off-white solid and used directly in the nextstep. LCMS-ESI (m/z) calculated for C₂₇H₃₃N₃O₅: 479.2. found 502.2[M+Na]⁺, t_(R)=4.11 min. ¹H NMR (400 MHz, CDCl₃) δ 7.91 (d, J=7.6 Hz,1H), 7.39 (d, J=7.5 Hz, 1H), 7.27 (t, J=7.7 Hz, 1H), 6.81 (ddd, J=20.2,12.8, 5.1 Hz, 3H), 5.18 (d, J=8.4 Hz, 1H), 4.69 (d, J=8.3 Hz, 1H), 4.15(d, J=6.1 Hz, 2H), 4.06-3.93 (m, 4H), 3.32 (ddd, J=17.4, 8.8, 3.4 Hz,1H), 3.14-2.91 (m, 1H), 2.65-2.40 (m, 1H), 1.75 (dq, J=12.9, 8.4 Hz,1H), 1.36 (ddd, J=40.2, 26.9, 22.6 Hz, 15H).

(S)-tert-butyl4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-57 was prepared in an analogous fashion from (R)-tert-butyl4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-ylcarbamate INT-55.

(R)-4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-aminehydrochloride (Compound 65)

To (R)-tert-butyl4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-56 (150 mg, 0.312 mmol) in dioxane (1 mL) was added 4N HCl indioxane (1 mL). The mixture was stirred at room temperature for 6 h, andproduct precipitated. The reaction mixture was diluted with Et₂O and thesolid collected by filtration to produce of(R)-4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-aminehydrochloride 65 (125 mg, 96%) as an off-white solid. LCMS-ESI (m/z):calcd for C₂₂H₂₅N₃O₃: 379.2. found 402.1 [M+Na]⁺, t_(R)=2.38 min. ¹H NMR(400 MHz, DMSO) δ 8.40 (s, 1H), 8.01 (d, J=7.6 Hz, 1H), 7.78 (d, J=7.2Hz, 1H), 7.53 (t, J=7.7 Hz, 1H), 7.02 (d, J=2.0 Hz, 1H), 6.90 (dt,J=8.2, 5.1 Hz, 2H), 4.81 (s, 1H), 4.35 (s, 2H), 4.01 (p, J=6.9 Hz, 4H),3.36 (s, 2H), 3.22-3.04 (m, 1H), 2.55-2.43 (m, 2H), 2.05 (dd, J=14.0,8.4 Hz, 1H), 1.32 (td, J=7.0, 4.0 Hz, 6H).

(S)-4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-aminehydrochloride INT-58 can be prepared in an analogous fashion from(S)-tert-butyl4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylcarbamateINT-57.

(R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-cyano-2,3-dihydro-1H-inden-1-yl)carbamate(INT-59)

To (R)-tert-butyl 4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate INT-52(0.700 g, 2.7 mmol) was added anhydrous DMF (10 mL) and the reactionmixture was stirred in a 0° C. ice bath under N₂. Sodium hydride (0.541g, 13.5 mmol) was added and the mixture was stirred at 0° C. for 2 h.After 2 h, (2-bromoethoxy)-tert-butyldimethylsilane (1.43 g, 5.9 mmol)was added and the reaction mixture was allowed to warm to roomtemperature for 1 h. The reaction was cooled to 0° C. and quenched withMeOH followed by saturated NaHCO₃. The mixture was extracted with EA andbrine. The combined organic layers were dried over MgSO₄, filtered, andconcentrated to produce a brown oil. The crude product was purified bysilica gel flash chromatography (20% EA/Hexanes) to afford 0.868 g (77%)of (R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-cyano-2,3-dihydro-1H-inden-1-yl)carbamateINT-59 as a yellow oil. LCMS-ESI (m/z) calculated for C₂₃H₃₆N₂O₃Si:416.6. found 317.1 [M+H−Boc]⁺, t_(R)=4.05 min. ¹H NMR (400 MHz,(CD₃)₂SO) δ 7.50 (m, 1H), 7.37 (m, 1H), 7.26 (m, 1H), 5.78 (m, 1H), 4.02(m, 2H), 3.51 (m, 2H), 3.29 (m, 1H), 2.97 (m, 1H), 2.26 (m, 2H), 1.40(s, 9H), 0.83 (s, 9H), 0.09 (s, 6H).

(S)-tert-butyl(4-cyano-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamate(INT-60)

(S)-tert-butyl(4-cyano-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamateINT-60 was prepared analogously to INT-59 from (S)-tert-butyl4-cyano-2,3-dihydro-1H-inden-1-ylcarbamate INT-53 and2-chloro-N,N-dimethylacetamide.

(R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(N-hydroxy-carbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamate(INT-61)

Prepared using General Procedure 1. To (R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-cyano-2,3-dihydro-1H-inden-1-yl)carbamate INT-59 (0.800g, 1.9 mmol) in EtOH (8 mL) was added hydroxylamine hydrochloride (0.400g, 5.8 mmol) and Na₂CO₃ (0.610 g, 5.8) and the reaction mixture washeated at 85° C. for 12 h. Once cooled to room temperature, the reactionmixture was filtered using EtOH to rinse the filter cake. The filtratewas concentrated under reduced pressure and washed with EA and brine.The combined organic layers were dried over MgSO₄, filtered, andconcentrated to produce 0.860 g (100%) of (R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-61 as a light yellow oil. LCMS-ESI (m/z) calculated forC₂₃H₃₉N₃O₄Si: 449.7. found 350.2 [M+H-Boc]⁺, t_(R)=1.97 min.

(S)-tert-butyl(2-(dimethylamino)-2-oxoethyl)(4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamate(INT-62)

(S)-tert-butyl(2-(dimethylamino)-2-oxoethyl)(4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-62 was prepared from (S)-tert-butyl(4-cyano-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamateINT-60 using General Procedure 1 and in an analogous fashion to INT-61.

(R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(5-(4-phenyl-5-(trifluoromethyl)thiophen-2-yl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)carbamate(INT-63)

Prepared using General Procedure 2. To a solution of4-phenyl-5-(trifluoromethyl)thiophene-2-carboxylic acid (0.109 g, 0.4mmol) in DMF (3.0 mL) was added HOBt (0.088 g, 0.57 mmol) and EDC (0.109g, 0.57 mmol) at room temperature. The reaction mixture was stirred for0.5 h until the complete formation of the HOBt-acid complex.(R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-61 (0.200 g, 0.44 mmol) was added and the mixture was stirred atroom temperature for 0.5 h until the formation of the intermediate(R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(N-(4-phenyl-5-(trifluoromethyl)thiophene-2-carbonyloxy)carbamimidoyl)-2,3-dihydro-1H-inden-1-yl) carbamate was observed. Thereaction mixture was heated at 85° C. for 4 h. Upon cooling, the mixturewas extracted with DCM and brine. The combined organic layers were driedover MgSO₄, filtered, and concentrated under reduced pressure to producea brown oil. The crude product was purified by silica gel flashchromatography (MeOH/DCM) to yield 0.108 g (40%) of (R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(5-(4-phenyl-5-(trifluoromethyl)thiophen-2-yl)-1,2,4-oxa-diazol-3-yl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-63 as a light yellow oil. LCMS-ESI (m/z) calculated forC₃₅H₄₂F₃N₃O₄SSi: 685.9. found 411.0 [M+H− tert-butyl2-(tert-butyldimethylsilyloxy)ethylcarbamate]⁺, t_(R)=4.01 min.

(S)-tert-butyl(4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamate(INT-64)

(S)-tert-butyl(4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamateINT-64 was prepared using General Procedure 2, analogously to INT-63,from (S)-tert-butyl(2-(dimethylamino)-2-oxoethyl)(4-(N-hydroxycarbamimidoyl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-62 and 2-(3,4-diethoxyphenyl)acetic acid.

(R)-2-(4-(5-(4-phenyl-5-(trifluoromethyl)thiophen-2-yl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylamino)ethanol(Compound 67)

To (R)-tert-butyl2-(tert-butyldimethylsilyloxy)ethyl(4-(5-(4-phenyl-5-(trifluoromethyl)thiophen-2-yl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)carbamateINT-63 (0.108 g, 0.16 mmol) dissolved in DCM (1.5 mL) was added 2N HClin ether (1.45 mL, 2.9 mmol). The solution was stirred at roomtemperature for 12 h. The solvent was removed under a stream of nitrogenand the product dried under vacuum to afford 0.052 g (65%) of(R)-2-(4-(5-(4-phenyl-5-(trifluoromethyl)thiophen-2-yl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-ylamino)ethanol 67 as the HCl salt. LCMS-ESI (m/z): calcd for C₂₄H₂₀F₃N₃O₂S:471.5. found 472.1 [M+H]⁺, t_(R)=7.43 min (Method 2). ¹H NMR (400 MHz,CDCl₃) δ 9.62 (s, 1H), 8.19 (d, J=7.6, 1H), 8.03 (d, J=7.5, 1H), 7.87(t, J=1.5, 1H), 7.53-7.40 (m, 6H), 4.86 (d, J=4.8, 1H), 3.88 (s, 2H),3.74-3.50 (m, 1H), 3.41 (ddd, J=13.3, 9.4, 4.4, 1H), 3.06 (m, 1H), 2.98(m, 1H), 2.67-2.42 (m, 2H). ¹³C NMR (100 MHz, DMSO) δ 169.22, 168.07,145.68, 144.75, 139.39, 135.43, 132.42, 129.42, 129.37, 129.25, 128.69,128.27, 127.62, 126.41, 123.16, 122.37, 120.47, 61.10, 56.63, 46.54,31.66, 27.80.

(S)-2-((4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)-N,N-dimethylacetamide(Compound 66)

(S)-2-((4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)amino)-N,N-dimethylacetamide66 was prepared analogously to compound 67 from (S)-tert-butyl(4-(5-(3,4-diethoxybenzyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl)(2-(dimethylamino)-2-oxoethyl)carbamateINT-64.

tert-butyl 5-cyano-1H-indole-1-carboxylate (INT-65)

To a flask containing 5-cyanoindole (500 mg, 3.52 mmol) in CH₃CN (5 mL)was added Boc₂O (920 mg, 4.22 mmol) and DMAP (42 mg, 0.35 mmol) and themixture was stirred at room temperature for 0.5 h. The mixture wasconcentrated, redissolved in DCM and chromatographed (EtOAc/hexanes) toprovide 766 mg (90%) of tert-butyl 5-cyano-1H-indole-1-carboxylateINT-65 as a white solid. LCMS-ESI (m/z) calculated for C₁₄H₁₄N₂O₂:242.27. found 243.1 [M+H]⁺, t_(R)=3.93 min.

tert-butyl 5-(N-hydroxycarbamimidoyl)-1H-indole-1-carboxylate (INT-66)

Prepared using General Procedure 1. To a flask containing tert-butyl5-cyano-1H-indole-1-carboxylate INT-65 (200 mg, 0.73 mmol) was addedEtOH (6 mL), hydroxylamine hydrochloride (177 mg, 2.54 mmol) and Na₂CO₃(154 mg, 1.45 mmol). The mixture was stirred at 75° C. overnight thenconcentrated, re-dissolved in DCM and washed with NaHCO₃. The combinedorganic layers were dried over Na₂SO₄ and concentrated to provide 222 mgof crude tert-butyl 5-(N-hydroxycarbamimidoyl)-1H-indole-1-carboxylateINT-66 as a white solid which was used directly in the next experiment.LCMS-ESI (m/z) calculated for C₁₄H₁₇N₃O₃: 275.3. found 276.1 [M+H]⁺,t_(R)=2.25 min.

tert-butyl5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1H-indole-1-carboxylate(INT-67)

Prepared using General Procedure 2. A flask containing3-cyano-4-isopropoxybenzoic acid (135 mg, 0.66 mmol), HOBt (130 mg, 0.85mmol) and EDC (164 mg, 0.85 mmol) in DMF (2.5 mL) was stirred for 1.5 hat room temperature under an atmosphere of N₂. A solution of crudetert-butyl 5-(N-hydroxycarbamimidoyl)-1H-indole-1-carboxylate INT-66(199 mg, 0.72 mmol) in DMF (2.5 mL) was added to the mixture. After 1 hat room temperature, the mixture was heated to 75° C. and stirredovernight. The reaction mixture was diluted with NaHCO₃ and extractedwith EtOAc. The combined organic extracts were dried over Na₂SO₄ andconcentrated. The resulting crude material was chromatographed(EtOAc/hexanes) to provide 174 mg (59%) of tert-butyl5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1H-indole-1-carboxylateINT-67 as a white solid. LCMS-ESI (m/z) calculated for C₂₅H₂₄N₄O₄:444.5. found 445.1 [M+H]⁺, t_(R)=3.67 min (Method 1).

5-(3-(1H-indol-5-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile(Compound 68)

To a flask containing tert-butyl5-(5-(3-cyano-4-isopropoxyphenyl)-1,2,4-oxadiazol-3-yl)-1H-indole-1-carboxylateINT-67 (75 mg, 0.17 mmol) was added dioxane (2 mL) followed by 4N HCl indioxane (0.5 mL, 2 mmol). The reaction mixture was stirred overnight atroom temperature then heated at 50° C. overnight. Additional 4NHCl/dioxane (0.5 mL, 2 mmol) was added and the mixture was heated at 50°C. for an additional 2 h to complete the deprotection. The reactionmixture was diluted with EtOAc and washed with NaHCO₃. The combinedorganic layers were washed with brine, dried over Na₂SO₄, andconcentrated. The material was purified by chromatography(EtOAc/hexanes) to provide 17 mg (30%) of5-(3-(1H-indol-5-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile 68as a white solid. LCMS-ESI (m/z) calculated for C₂₀H₁₆N₄O₂: 344.5. found345.1 [M+H]⁺, t_(R)=2.34 min (Method 1). ¹H NMR (400 MHz, CDCl₃) δ8.51-8.47 (m, 1H), 8.45 (d, J=2.2 Hz, 1H), 8.38 (d, J=7.2 Hz, 1H), 8.35(dd, J=8.9, 2.2 Hz, 1H), 7.99 (dd, J=8.5, 1.6 Hz, 1H), 7.51 (d, J=8.5Hz, 1H), 7.33-7.28 (m, 1H), 7.11 (d, J=9.0 Hz, 1H), 6.72-6.64 (m, 1H),4.79 (dt, J=12.2, 6.1 Hz, 1H), 1.47 (t, J=5.8 Hz, 6H).

N-hydroxybenzofuran-5-carboximidamide (INT-68)

Prepared using General Procedure 1. To a flask containingbenzofuran-5-carbonitrile (200 mg, 0.73 mmol) was added EtOH (6 mL),hydroxylamine hydrochloride (176.7 mg, 2.54 mmol) and Na₂CO₃ (154 mg,1.42 mmol). The mixture was stirred at 75° C. overnight thenconcentrated, re-dissolved in DCM and washed with NaHCO₃. The combinedorganic layers were dried over Na₂SO₄, and concentrated to provide 222mg of crude N-hydroxybenzofuran-5-carboximidamide INT-68 as a whitesolid which was used directly in the next step without purification.LCMS-ESI (m/z) calculated for C₉H₈N₂O₂: 176.2. found 177.1 [M+H]⁺,t_(R)=0.83 min.

5-(3-(benzofuran-5-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile(Compound 69)

Prepared using General Procedure 2. A flask containing3-cyano-4-isopropoxybenzoic acid (147.7 mg, 0.72 mmol), HOBt (143 mg,0.94 mmol) and EDC (180 mg, 0.94 mmol) in DMF (2.0 mL) was stirred for0.5 h at room temperature under an atmosphere of N₂. A solution ofN-hydroxybenzofuran-5-carboximidamide INT-68 (218 mg, 0.79 mmol) in DMF(2.0 mL) was added to the mixture. After 1 h at room temperature, themixture was stirred at 85° C. overnight. The reaction mixture wasdiluted with NaHCO₃ and extracted with EA. The combined organic extractswere dried over Na₂SO₄, and concentrated. The resulting crude materialwas chromatographed (EA/hexanes) to provide 110 mg (44%) of5-(3-(benzofuran-5-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile 69as a white solid. LCMS-ESI (m/z) calculated for C₂₀H₁₅N₃O₃: 345.4. found346.1 [M+H]⁺, t_(R)=2.77 min (Method 1). ¹H NMR (400 MHz, CDCl₃) δ 8.45(dd, J=4.5, 1.9 Hz, 2H), 8.35 (dd, J=8.9, 2.2 Hz, 1H), 8.12 (dd, J=8.6,1.7 Hz, 1H), 7.71 (d, J=2.2 Hz, 1H), 7.63 (d, J=8.6 Hz, 1H), 7.13 (d,J=9.0 Hz, 1H), 6.88 (dd, J=2.2, 0.8 Hz, 1H), 4.80 (s, 1H), 1.48 (d,J=6.1 Hz, 6H).

N-hydroxy-3-methylisonicotinimidamide (INT-69)

Prepared using General Procedure 1. To 3-methylisonicotinonitrile (0.500g, 4.2 mmol) in EtOH (7 mL) was added hydroxylamine hydrochloride (0.588g, 8.5 mmol) and Na₂CO₃ (1.34 g, 12.7 mmol) and the reaction mixture washeated at 85° C. for 4 h. Once cooled to room temperature, the reactionmixture was filtered using EtOH to rinse the filter cake. The filtratewas concentrated under reduced pressure. The resulting pale yellow solidwas triturated with ice water (50 mL), filtered, and the solid waswashed with ice water (5 mL). The solid was dried under reduced pressureto yield 0.47 g (74%) of N-hydroxy-3-methylisonicotinimidamide INT-69 asa white powder. LCMS-ESI (m/z) calculated for C₇H₉N₃O: 151.2. found152.1 [M+H]⁺, t_(R)=0.56 min. ¹H NMR (400 MHz, CD₃OD) δ 8.43-8.32 (m,2H), 7.34 (d, J=5.0, 1H), 2.39 (s, 3H).

2-isopropoxy-5-(3-(3-methylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)benzonitrile(Compound 70)

Prepared using General Procedure 2. To a solution of3-cyano-4-isopropoxybenzoic acid (0.122 g, 0.60 mmol) in DMF (1.5 mL)was added HOBt (0.132 g, 0.86 mmol) and EDC (0.165 g, 0.86 mmol) at roomtemperature. The reaction was stirred for 0.5 h until the completeformation of the HOBt-acid complex.N-hydroxy-3-methylisonicotinimidamide INT-69 (0.100 g, 0.66 mmol) wasadded and the mixture was stirred at room temperature for 0.5 h untilformation of the intermediateN-(3-cyano-4-isopropoxybenzoyloxy)-3-methylisonicotinimidamide wasobserved. The reaction mixture was then heated at 80° C. for 4 h. Uponcooling, the mixture was extracted with DCM and brine. The combinedorganic layers were dried over MgSO₄, filtered, and concentrated underreduced pressure to produce a brown oil. The crude productrecrystallized from MeOH (3 mL) and the resulting crystals were filteredand washed with cold MeOH to yield product as a white crystalline solid.To the product was added Et₂O (0.5 mL) followed by 2N HCl in Et₂O (0.6mL). The mixture was stirred at room temperature for 10 minutes thendried under nitrogen and subsequently under vacuum to afford 0.087 g(45%) of2-isopropoxy-5-(3-(3-methylpyridin-4-yl)-1,2,4-oxadiazol-5-yl)benzonitrile70 as the HCl salt. LCMS-ESI (m/z) calculated for C₁₈H₁₆N₄O₂: 320.3.found 321.1 [M+H]⁺, t_(R)=8.82 min (Method 2). ¹H NMR (400 MHz, CDCl₃) δ8.80 (d, J=17.5, 2H), 8.70 (d, J=5.7, 1H), 8.44 (d, J=2.2, 1H), 8.36(dd, J=8.9, 2.2, 1H), 7.18 (d, J=9.1, 1H), 4.83 (dt, J=12.2, 6.1, 1H),2.95 (s, 3H), 1.49 (d, J=6.1, 6H). ¹³C NMR (101 MHz, DMSO) δ 173.90,166.58, 162.81, 147.11, 142.99, 137.55, 135.01, 134.79, 134.06, 125.00,115.42, 115.17, 115.00, 102.57, 72.66, 21.48, 18.69.

4-bromo-2-((tert-butyldimethylsilyloxy)methyl)pyridine (INT-70)

To a stirring solution of (4-bromopyridin-2-yl)methanol (1.50 g, 8.0mmol) in DCM (4 mL) was added tert-butylchlorodimethylsilane (1.20 g,8.0 mmol) following by TEA (1.60 g, 12.0 mmol). The reaction mixture wasstirred at room temperature for 12 h then washed with brine and EA. Thecombined organic layers were dried over MgSO₄, filtered, andconcentrated to yield an amber liquid. The crude product was purified bychromatography (EA/Hexanes) to produce 1.67 g (70%) of4-bromo-2-((tert-butyldimethylsilyloxy)methyl)pyridine INT-70 as a lightyellow liquid. LCMS-ESI (m/z) calculated for C₁₂H₂₀BrNOSi: 302.3. found303.0 [M+H]⁺, t_(R)=4.87 min (Method 1). ¹H NMR (400 MHz, CDCl₃) δ 8.30(d, J=5.3, 1H), 7.68 (dd, J=1.9, 0.7, 1H), 7.35-7.24 (m, 1H), 4.80 (s,2H), 0.99-0.86 (m, 9H), 0.16-0.06 (m, 6H).

2-((tert-butyldimethylsilyloxy)methyl)isonicotinonitrile (INT-71)

To a stirring solution of4-bromo-2-((tert-butyldimethylsilyloxy)methyl)pyridine INT-70 (0.800 g,2.6 mmol) in 3 mL of 1-methy-2-pyrrolidine (NMP) was added zinc cyanide(0.610 g, 5.2 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.060g, 0.052 mmol). The solution was degassed with N₂ and the reactionmixture heated at 95° C. for 12 h. Upon cooling, the reaction mixturewas diluted with saturated NaHCO₃ and extracted with DCM. The combinedorganic layers were dried over MgSO₄, filtered, and concentrated. Thecrude product was purified by chromatography (MeOH/DCM) to produce 0.170g (26%) of 2-((tert-butyldimethylsilyloxy)methyl)isonicotinonitrileINT-71 as a light yellow solid. LCMS-ESI (m/z) calculated forC₁₃H₂₀N₂OSi: 248.4. found 249.1 [M+H]⁺, t_(R)=4.21 min (Method 1).

2-((tert-butyldimethylsilyloxy)methyl)-N-hydroxyisonicotinimidamide(INT-72)

Prepared using General Procedure 1. To2-((tert-butyldimethylsilyloxy)methyl)isonicotinonitrile INT-71 (0.169g, 0.68 mmol) in EtOH (8 mL) was added hydroxylamine hydrochloride(0.142 g, 2.0 mmol) and Na₂CO₃ (0.216 g, 2.0 mmol) and the reactionmixture was heated at 85° C. for 12 h. Once cooled to room temperature,the reaction mixture was filtered using EtOH to rinse the filter cake.The filtrate was concentrated under reduced pressure and washed with EAand brine. The combined organic layers were dried over MgSO₄, filtered,and concentrated to produce 0.191 g (100%) of2-((tert-butyldimethylsilyloxy)methyl)-N-hydroxyisonicotinimidamideINT-72 as a light yellow oil. LCMS-ESI (m/z) calculated forC₁₃H₂₃N₃O₂Si: 281.4. found 282.1 [M+H]⁺, t_(R)=2.76 min (Method 1). ¹HNMR (400 MHz, (CD₃)₂SO) δ 8.50 (dd, J=5.2, 0.7, 1H), 7.74 (dd, J=1.6,0.7, 1H), 7.51 (dd, J=5.2, 1.7, 1H), 5.98 (s, 2H), 0.96-0.89 (m, 9H),0.14-0.07 (m, 6H).

5-(3-(2-(hydroxymethyl)pyridin-4-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxy-benzonitrile(Compound 71)

Prepared using General Procedure 2. To a solution of3-cyano-4-isopropoxybenzoic acid (0.033 g, 0.16 mmol) in DMF (1.0 mL)was added HOBt (0.036 g, 0.23 mmol) and EDC (0.045 g, 0.23 mmol) at roomtemperature. The reaction was stirred for 0.5 h until the completeformation of the HOBt-acid complex.2-((Tert-butyldimethylsilyloxy)methyl)-N-hydroxyisonicotinimidamideINT-72 (0.050 g, 0.18 mmol) was added and the mixture was stirred atroom temperature for 0.5 h until the formation of the intermediate2-((tert-butyldimethylsilyloxy)methyl)-N-(3-cyano-4-isopropoxybenzoyloxy) isonicotinimidamide was observed. The reaction mixture wasthen heated at 85° C. for 4 h. To the cooled reaction mixture MeOH (1.0mL) was added, and the solution was filtered. The resulting filtrate waspurified by preparative HPLC to produce 5.6 mg (8%) of5-(3-(2-(hydroxymethyl)pyridin-4-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile71 as the TFA salt. LCMS-ESI (m/z) calculated for C₁₈H₁₆N₄O₃: 336.3.found 337.1 [M+H]⁺, t_(R)=7.45 min (Method 2). ¹H NMR (400 MHz, CD₃OD) δ8.78 (d, J=5.5, 1H), 8.48 (dd, J=10.6, 8.3, 1.4, 3H), 8.23 (dd, J=5.5,1.6, 1H), 7.48 (d, J=9.0, 1H), 4.93 (s, 2H), 4.89 (m, 1H), 1.48 (d,J=6.1, 6H).

Selected compounds and their corresponding analytical data is shown inTable 1, where the LCMS data was collected using Method 2 (see GeneralMethods). The enantiomeric purity was determined for key intermediatesand selected final compounds and is presumed from the synthesis for theremaining compounds.

TABLE 1 LCMS RETEN- COMPOUND TION STRUCTURE NUMBER TIME (min)

1 9.32

2 9.32

3 6.35

4 6.34

5 9.21

6 9.20

7 8.09

8 8.08

9 8.25

10 8.26

11 9.53

12 9.53

13 8.16

14 8.16

15 9.01

16 9.03

17 8.55

18 8.56

19 8.31

20 6.45

21 8.90

22 8.89

23 9.37

24 9.36

25 6.56

26 8.82

27 8.8

28 9.41

29 9.36

30 9.87

31 9.83

32 9.68

33 9.66

34 8.83

35 8.84

36 8.73

37 8.76

38 8.47

39 8.49

40 9.09

41 9.07

42 9.00

43 9.02

44 6.74

45 6.82

46 6.69

47 6.58

48 6.55

49 6.54

50 6.36

51 6.40

52 6.13

53 6.52

54 6.71

55 6.76

56 8.63

57 6.16

58 6.34

59 5.85

60 8.56

61 6.07

62 6.22

63 6.33

64 6.43

65 6.00

66 6.23

67 7.40

68 9.66

69 10.74

70 8.81

71 7.44

Biological Assays Assay Procedures

Generation of S1P₁-Mediated Inhibition of cAMP Reporter Assay

A mammalian expression plasmid containing S1P₁/EDG1 cloned into pcDNA3.1was purchased from Missouri S&T cDNA Resource Centre. The nucleotide andamino acid sequence of human S1P₁/EDG1 are published in Hla and Maciag(J Biol Chem, 265(1990), 9308-9313). S1P₁/pcDNA3.1 was transfected intothe CRE-bla CHO K1 (Invitrogen) cell line, and stable single cell cloneswere selected using standard techniques. Expression of functionalS1P₁/EDG1 receptor was confirmed by cell surface FACS with a S1P₁antibody (R&D Systems, clone 218713) and S1P-mediated inhibition ofForskolin induced cAMP.

S1P₁ CRE-Bla CHOK1 Reporter Assay—Characterization of S1P¹ Agonists

Cells were seeded into 384-well black wall/clear bottom plates at 10⁴cells/well/19.5 μl assay media (DMEM-phenol free, 0.5% charcoal/dextranstripped serum, 2 mM glutamine, 0.1 mM NEAA, 1 mM Na-Pyruvate, 25 mMHepes) and incubated for 18 hrs at 37° C. in 5% CO₂. Dose responsecurves (10-point) were generated in 10 mM Hepes, 0.1% Pluronic F127, inthe presence of Forskolin. Cells were treated with 0.5 μl compound inthe presence of 2 μM Forskolin for 4 hrs at 37° C. The FRET-basedβ-lactamase fluorescent substrate (LiveBLAzer™-FRET B/G Loading KitCC4-AM; Invitrogen) was prepared according to manufacturer's directions,and incubated with cells for 2 hrs at room temperature. Plates were readat Ex:410/Em:458 and Ex:410/Em:522, and the response ratio determined.Data was analyzed by non-linear regression to determine the EC50 forinhibition of Forskolin induced cAMP.

Specificity Over Other S1P Receptors

To assess compound specificity on other S1P receptors the following celllines were used: S1P₂ CRE-bla CHOK1, S1P₃-Gα15 NFAT-bla HEK293T(Invitrogen), S1P₄-bla TANGO U2OS (Invitrogen), S1P₅-bla TANGO U2OS(Invitrogen). The same assay set up for S1P₁ was used but withoutForskolin. S1P₄ and S1P₅ assays were performed in FreeStyle Expressionmedium (Invitrogen). S1P₅ cells were incubated for 48 hrs in prior totreatment with compound.

Reported S1P₁ Activity

Activity data for selected S1P₁ agonists is displayed in Table 2. Theactivity range is denoted as follows: ++++ denotes agonist activity<0.05 nM. +++ denotes agonist activity between 0.05 to 0.50 nM, and ++denotes agonist activity between 0.50-5.00 nM, and + denotes agonistactivity >5.00 nM. N/A denotes not available.

TABLE 2 COMPOUND S1P₁ NUMBER ACTIVITY 1 +++ 2 ++++ 3 ++ 4 +++ 5 +++ 6+++ 7 +++ 8 +++ 9 +++ 10 +++ 11 +++ 12 +++ 13 +++ 14 +++ 15 +++ 16 ++ 17++ 18 ++ 19 ++ 20 ++ 21 +++ 22 +++ 23 ++ 24 +++ 25 ++ 26 +++ 27 ++++ 28+++ 29 +++ 30 +++ 31 ++ 32 +++ 33 +++ 34 ++++ 35 ++++ 36 ++++ 37 ++++ 38++++ 39 ++++ 40 ++++ 41 ++++ 42 ++++ 43 ++++ 44 +++ 45 ++ 46 ++ 47 +++48 +++ 49 +++ 50 +++ 51 +++ 52 ++++ 53 +++ 54 ++ 55 +++ 56 ++ 57 ++ 58 +59 + 60 + 61 + 62 + 63 + 64 + 65 + 66 + 67 + 68 ++ 69 ++ 70 ++++ 71 +++

S1P₁-S1P₅ data for specific compounds is presented in Table 3. Theagonist values (EC₅₀) are reported in nM.

TABLE 3 COMPOUND NUMBER S1P₁ S1P₂ S1P₃ S1P₄ S1P₅ 80.143 >10000 >10000 >10000 108.9 13 0.100 >10000 >10000 >10000 77.0 290.065 >10000 >10000 >10000 37.8 33 0.192 >10000 >10000 616.7 260.1 370.024 1437 >10000 879.4 3.5 49 0.104 >10000 >10000 >10000 94.6

In Vivo Assays Determination of Absolute Oral Bioavailability in Rats.

Pharmacokinetic studies are conducted in non-fasted femaleSprague-Dawely rats (Simonsen Laboratories or Harlan Laboratories). Ratsare housed in an ALAAC accredited facility and the research is approvedby the facilities Institutional Animal Care and Use Committee (IACUC).The animals are acclimated to the laboratory for at least 48 h prior toinitiation of experiments.

Compounds are formulated in 5% DMSO/5% Tween20 and 90% purified water(intravenous infusion) or 5% DMSO/5% Tween20 and 90% 0.1N HCL (oralgavage). The concentration of the dosing solutions is verified byHPLC-UV. For intravenous dosing, compounds are administered by aninfusion pump into the jugular vein over one minute to manuallyrestrained animals (n=4 rats/compound). Oral dosing is by gavage using astandard stainless steel gavage needle (n=2-4 rats/compound). For bothroutes of administration, blood is collected at eight time-points afterdosing with the final sample drawn 24 h post dose. Aliquots of the bloodsamples are transferred to polypropylene 96-well plate and frozen at−20° C. until analysis.

After thawing the blood samples at room temperature, 54 of DMSO is addedto each well. Proteins are precipitated by adding 150 μL acetonitrilecontaining 200 nM internal standard(4-hydroxy-3-(alpha-iminobenzyl)-1-methyl-6-phenylpyrindin-2-(1H)-one)and 0.1% formic acid. Plates are mixed for 1 min on a plate shaker tofacilitate protein precipitation and then centrifuged at 3,000 rpm for10 min to pellet protein. The supernatant is transferred to a cleanplate and centrifuged at 3,000 rpm for 10 min to pellet any remainingsolid material prior to LC/MS/MS analysis. Calibration curve standardsare prepared by spiking 54 compound stock in DMSO into freshly collectedEDTA rat blood. An eight point standard curve spanning a range of 5 nMto 10,000 nM is included with each bio-analytical run. The standards areprocessed identically to the rat pharmacokinetic samples.

Concentrations in the rat pharmacokinetic samples are determined using astandardized HPLC-LC/MS/MS method relative to the eight point standardcurve. The system consists of a Leap CTC Pal injector, Agilent 1200 HPLCwith binary pump coupled with an Applied Biosystems 3200 QTrap.Compounds are chromatographed on a Phenomenex Synergy Fusion RP 20×2 mm2 um Mercury Cartridge with Security Guard. A gradient method is usedwith mobile phase A consisting of 0.1% formic acid in water and mobilephase B consisting of 0.1% formic acid in acetonitrile at flow ratesvarying from 0.7 to 0.8 mL/min. Ions are generated in positiveionization mode using an electrospray ionization (ESI) interface.Multiple reaction monitoring (MRM) methods are developed specific toeach compound. The heated nebulizer is set at 325° C. with a nebulizercurrent of 4.8 μA. Collision energies used to generate daughter ionsrange between 29 and 39 V. Peak area ratios obtained from MRM of themass transitions specific for each compound are used for quantification.The limit of quantification of the method is typically 5 nM. Data arecollected and analyzed using Analyst software version 1.4.2.

Blood and/or plasma concentration versus time data are analyzed usingnon-compartmental methods (WinNonlin version 5.2; model 200 for oraldosing and model 202 for intravenous infusion). Absolute oralbioavailability (%) is calculated using the following expression: (OralAUC×IV Dose)/(IV AUC×Oral Dose)×100.

Lymphopenia

In mice: Female C57BL6 mice (Simonsen Laboratories, Gilroy Calif.) arehoused in an ALAAC accredited facility and the research is approved bythe facilities Institutional Animal Care and Use Committee (IACUC). Theanimals are acclimated to the laboratory for at least 5 days prior toinitiation of experiments. Mice (n=3/compound/time-point) are dosed byoral gavage with 1 mg/kg compound formulated in a vehicle consisting of5% DMSO/5% Tween 20 and 90% 0.1N HCl. Control mice are dosed PO with thevehicle. Terminal whole blood samples are collected from isofluraneanesthetized mice by cardiac puncture into EDTA. Whole blood isincubated with rat anti-mouse CD16/CD32 (Mouse BD Fc Block, #553141),PE-Rat anti-mouse CD45R/B220 (BD #553089), APC-Cy7-Rat anti-mouse CD8a(BD #557654), and Alexa Fluor647-Rat anti-mouse CD4 (BD #557681) for 30min on ice. Red blood cells are lysed using BD Pharm Lyse Lysing buffer(#555899) and white blood cells are analyzed by FACS. Lymphopenia isexpressed as the % of white blood cells that are CD4 or CD8 positive Tcells. The overall lymphopenia response over 24 h is estimated bycalculating the area under the effect curve (AUEC) using the lineartrapezoidal rule.

In rats: Female rats (Simonsen Laboratories, Gilroy Calif.) are housedin an ALAAC accredited facility and the research is approved by thefacilities Institutional Animal Care and Use Committee (IACUC). Theanimals are acclimated to the laboratory for at least 5 days prior toinitiation of experiments. Rats (n=3/compound/time-point) are dosed byoral gavage with 1 mg/kg compound formulated in a vehicle consisting of5% DMSO/5% Tween 20 and 90% 0.1N HCL. Control rats are dosed PO with thevehicle. Whole blood is collected from isoflurane anesthetized rats viathe retro-orbital sinus and terminal samples are collected by cardiacpuncture into EDTA. Whole blood is incubated with mouse anti-rat CD32(BD #550271), PE-mouse anti-rat CD45R/B220 (BD #554881), PECy5-mouseanti-rat CD4 (BD #554839), and APC-mouse anti-rat CD8a (eBioscience#17-0084) for 30 minutes on ice. Red blood cells are lysed using BDPharm Lyse Lysing buffer (#555899) and white blood cells are analyzedwith a BD FACSArray. Lymphopenia is expressed as the % of white bloodcells that are CD4 or CD8 positive T cells. The overall lymphopeniaresponse over 24 h is estimated by calculating the area under the effectcurve (AUEC) using the linear trapezoidal rule. In some experiments,total lymphocyte counts are determined using a standard impediance basedveterinary hematology analyzer (IDEXX Preclinical Research Services,Sacramento, Calif.).

Evaluation of Therapeutic Index in Rats

Studies may be conducted in non-fasted male and female Sprague-Dawelyrats (Simonsen Laboratories). Rats may be housed in an AAALAC accreditedfacility and the research can be approved by the facilitiesInstitutional Animal Care and Use Committee (IACUC). The animals shouldbe acclimated to the laboratory for at least 5 days prior to initiationof experiments.

The compounds may be formulated as suspensions in a vehicle consistingof 0.5% carboxymethyl cellulose (Acros Organics) in purified water (pHadjusted to ˜2.2 with hydrochloric acid). The same formulation is usedin the rat lymphopenia and toxicology studies described below. Theconcentration of each compound in suspension should be verified to bewithin ±10% of the target concentration by HPLC-UV.

Prior to the conduct of toxicology studies, the effect of three to fivedaily doses of each compound on peripheral T-cell counts of female ratsmay be determined (see lymphopenia measurements in rats above). In theselymphopenia studies, blood samples are collected onto EDTA at intervalsafter the final study dose. The collection times need not be identicalfor each study; however, all studies may include a sample collected 24hours after the final dose. The lymphopenia data is used as a biomarkerto select equally pharmacologically active doses for the subsequenttoxicology study. The low dose for the toxicology study is the dose ofeach compound that resulted in a 50% reduction of T-cell count 24 hafter the final dose in the lymphopenia study relative to vehicletreated rats.

In the toxicology studies, three male and three female rats per groupare assigned to dosing groups using body weight based randomization. Acontrol group in each study receives vehicle. All animals are dosedorally by gavage on 5 or 14-consecutive days at a dose volume of 5mL/kg/day. The animals are observed daily for any manifestations ofadverse effect. Twenty-four hours after the final study dose, the ratsare anesthetized with isoflurane and a terminal blood sample is taken byintra-cardiac puncture for hematology and clinical chemistry evaluation(IDEXX Laboratories, Sacramento, Calif.). The lungs with trachea arecollected, weighed, and then prepared for histology by perfusion with10% neutral buffered formalin via the trachea. The internally fixedlungs are then preserved in 10% neutral buffered formalin and submittedfor histological examination (IDEXX).

The dose of each compound resulting in a 10% increase in the lung toterminal body weight ratio can be estimated for each compound by linearinterpolation. The therapeutic index can then be estimated as the ratioof the dose producing 10% lung weight increase to the dose producing 50%T-Cell depletion.

Description of the TNBS Crohn's Colitis Model in Rats

Male Sprague-Dawley rats (180-200 g) are acclimatized for seven days andthen assigned to 8 rats per group so that each group has approximatelythe same mean weight. Twenty-four hours prior to disease initiation,rats are deprived of food. Rats are anaesthetized and weighed, then 80mg/kg TNBS solution (50% TNBS: 50% 200 proof ethanol) is instilled intocolon via a 20 g feeding needle inserted into the anus. The rats aremaintained in head down position until recovery from anesthesia. Dailyoral dosing is initiated 2 h post TNBS-instillation for six days.Prednisolone serves as a positive control and is administered orallydaily at 10 mg/kg. Body weights are monitored daily and 24 h after thelast dose, all groups are terminated. The colon is removed, flushed offecal matter and examined for gross changes including strictures,adhesions and ulcers. The colon length, weight of the distal 2 cm, andwall thickness is recorded.

Description of Influenza A H1N1 Model in Mice

Male C57B1/6 (6-8 weeks of age) may be acclimatized for seven days andthen assigned to 5-8 mice per group so that each group has approximatelythe same mean weight. Mice may be infected with 10⁴ PFUs mouse-adaptedinfluenza A virus (A/WSN/33) via the intra-tracheal route. Mice may thenbe treated with 0.2-1.5 mg/kg compound p.o. 1 hr post-infection. Fortyeight hours after infection mice may be euthanized by cervicaldislocation and bronchoalveolar lavage fluid can be collected.Quantitative cytokine analysis may be performed via ELISA. In someexperiments whole body perfusion can be performed and lungs can becollected for cellular enumeration of inflammatory cells. Longevitystudies may be performed by infection with 3-10×10⁴ PFUs mouse-adaptedinfluenza A virus over 14 days.

1. A compound having the structure of Formula I-R or I-S or apharmaceutically acceptable salt, ester, prodrug, homolog, hydrate orsolvate thereof:

wherein X is —NR′R″ or —OR′″; Y is —CN, —Cl, or —CF₃; R′ is H, C₁₋₄alkyl, n-hydroxy C₁₋₄ alkyl, —SO₂—R¹, or —CO—R¹; R″ is H, —SO₂—R³, C₁₋₄alkyl optionally substituted with 1 or more R², or a ring moietyoptionally substituted with R⁴ wherein such ring moiety is piperidinyl,cyclohexyl, morpholinyl, thiazolyl, pyrazolyl, pyrrolidinyl, imidazolyl,or phenyl; or R′ and R″ taken together with the nitrogen atom to whichthey are bound form a 4, 5, or 6 membered saturated heterocyclic ringcontaining 0 or 1 additional heteroatoms where such additionalheteroatom is O or N wherein such heterocycle is optionally singly ormultiply substituted with substituents independently selected from thegroup consisting of —OH, oxo, —NH₂, n-hydroxy-C₁₋₄ alkyl, —COOH,—(CH₂)_(m)—COOH, —(CH₂)_(m)—COOR¹, —N(R¹R¹), and —(CH₂)_(m)—CO—N(R⁵R⁵);R′″ is H, C₁₋₄ alkyl, or —CO—R¹; each R¹ is independently C₁₋₄ alkyl orH; each R² is independently H, halo, OH, oxo, ═NH, NH₂, —COOH, F, —NHR¹,—N(R⁵R⁵),—SO₂—R¹, —SO₂—N(R⁵R⁵), —N(R¹)—SO₂—R¹, —COOR¹, —OCO—R¹,—CO—N(R⁵R⁵), —N(R¹)—COR¹, C₁₋₃ alkyl, C₁₋₃ alkoxy, and a ring moietyoptionally substituted with R⁴ wherein such ring moiety is piperazinyl,piperidinyl, morpholinyl, pyrrolidinyl, pyrazolyl, imidazolyl,benzimidazolyl, azetidinyl, cyclobutinyl, or phenyl; each R³ isindependently R², C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or C₁₋₄ alkyl optionallysubstituted with 1 or more R²; each R⁴ is independently halo, OH, —NH₂,—NHR¹, —N(R¹R¹), —COOH, —COOR¹, —NHCO—R¹, each R⁵ is independently C₁₋₄alkyl or H, or two R⁵ taken together with the nitrogen atom to whichthey are bound form a 4, 5, or 6 membered saturated heterocyclic ringcontaining 0 or 1 additional heteroatoms where such additionalheteroatom is O or N wherein such heterocycle is optionally substitutedwith —OH, —NH₂, —N(R¹R¹), n-hydroxy C₁₋₄ alkyl, —(CH₂)_(m)—COOH,—(CH₂)_(m)—COOR¹; and each m is independently 0, 1, 2, or
 3. 2. Thecompound of claim 1 wherein the compound has the structure of FormulaI-R or a pharmaceutically acceptable salt, ester, prodrug, homolog,hydrate or solvate thereof.
 3. The compound of claim 1 wherein thecompound has the structure of Formula I-S or a pharmaceuticallyacceptable salt, ester, prodrug, homolog, hydrate or solvate thereof. 4.The compound of claim 1 wherein the compound is substantiallyenantiomerically pure. 5-9. (canceled)
 10. The compound of claim 1wherein Y is Cl.
 11. The compound of claim 1 wherein Y is CF₃.
 12. Thecompound of claim 1 wherein Y is CN.
 13. The compound of claim 1 whereinX is —NR′R″.
 14. The compound of claim 1 wherein X is —OR′″.
 15. Thecompound of claim 14 wherein X is —OH.
 16. The compound of claim 14wherein X is —OCO—R¹.
 17. The compound of claim 16 wherein R¹ is C₁₋₃alkyl.
 18. The compound of claim 13 wherein R′ is H.
 19. The compound ofclaim 13 wherein R′ is —COR¹.
 20. The compound of claim 13 wherein R′ is—SO₂—R¹.
 21. The compound of claim 13 wherein R″ is H.
 22. The compoundof claim 13 wherein R″ is —SO₂—R³.
 23. The compound of claim 13 whereinR″ is C₁₋₄ alkyl optionally substituted with 1 or more R².
 24. Thecompound of claim 13 wherein R″ is —(CR^(a)R^(b))_(n)—R²; each R^(a) andeach R^(b) is independently selected from the group consisting of H,hydroxyl and methyl or R^(a) and R^(b) bound to the same carbon takentogether are oxo; and n is 0, 1, 2, or
 3. 25. The compound of claim 24wherein n is
 2. 26. The compound of claim 25 wherein R² is —OH, —NH₂,—NHR¹, —N(R⁵R⁵), or —COOH.
 27. The compound of claim 22 wherein R³ isC₁₋₄ alkyl optionally substituted with 1 or more R².
 28. The compound ofclaim 22 wherein Y is CN.
 29. The compound of claim 27 wherein R³ is—C₂H₅—N((R⁵R⁵) or —CH₂—CO—N(R⁵R⁵).
 30. The compound of claim 28 whereinR³ is C₂H₅—O—R¹.
 31. The compound of claim 12 wherein X is—NH—CO—N(R⁵R⁵).
 32. The compound of claim 1 wherein the compound isselected from the group consisting of:

or a pharmaceutically acceptable salt, ester, prodrug, homolog, hydrateor solvate thereof.
 33. The compound of claim 32 selected from the groupconsisting of:

or a pharmaceutically acceptable salt, ester, prodrug, homolog, hydrateor solvate thereof.
 34. A pharmaceutical composition comprising acompound of claim 1 and a suitable excipient.
 35. A pharmaceuticalcombination comprising a compound of claim 1 and a second medicament.36. The combination of claim 35 wherein the second medicament ismedically indicated for the treatment of multiple sclerosis, transplantrejection, or acute respiratory distress syndrome.
 37. (canceled)
 38. Amethod of activation or agonism of a sphingosine-1-phosphate receptorsubtype 1 comprising contacting the receptor subtype 1 with an effectiveamount of the compound of claim 1 or the composition of claim
 34. 39.The method of claim 38 wherein the compound activates or agonizes thesphingosine-1-phosphate receptor subtype 1 to a greater extent than thecompound activates or agonizes a sphingosine-1-phosphate receptorsubtype
 3. 40. The method of claim 38 wherein thesphingosine-1-phosphate receptor subtype 1 is disposed within a livingmammal.
 41. A method of treatment of a malcondition in a patient forwhich activation or agonism of an sphingosine-1-phosphate receptorsubtype 1 is medically indicated, comprising administering an effectiveamount of the compound of claim 1 to the patient at a frequency and fora duration of time sufficient to provide a beneficial effect to thepatient.
 42. The method of claim 41 wherein selective activation oragonism of an S1P subtype 1 receptor with respect to other subtypes ofS1P receptor is medically indicated.
 43. The method of claim 41 whereinthe malcondition comprises multiple sclerosis, transplant rejection,acute respiratory distress syndrome, ulcerative colitis, influenza,Crohn's disease or adult respiratory distress syndrome. 44-60.(canceled)